NASAL-SPRAY SAMPLING

Abstract

A method is provided that includes intranasally dispensing nasal wash fluid (26) into a nasal cavity (22) of a subject such that the nasal wash fluid (26) washes biological material into an oropharynx (30) of the subject from (a) the nasal cavity (22), (b) a nasopharynx (32) of the subject, or (c) the nasal cavity (22) and the nasopharynx (32). Thereafter, a specimen sample (24) is collected that passed out of an anterior opening (34) of an oral cavity (36) of the subject and contains at least a portion of the biological material washed into the oropharynx (30) by the nasal wash fluid (26). Other embodiments are also described.

Claims

1. A method comprising: intranasally dispensing nasal wash fluid into a nasal cavity of a subject such that the nasal wash fluid washes biological material into an oropharynx of the subject from (a) the nasal cavity, (b) a nasopharynx of the subject, or (c) the nasal cavity and the nasopharynx; and thereafter, collecting a specimen sample that passed out of an anterior opening of an oral cavity of the subject and contains at least a portion of the biological material washed into the oropharynx by the nasal wash fluid.

2. The method according to claim 1, wherein the biological material includes one or more particulates selected from the group consisting of: a virus, a bacterium, a protozoan, and a fungus.

3. The method according to claim 1, wherein intranasally dispensing comprises intranasally dispensing, by the subject, the nasal wash fluid into the nasal cavity.

4. The method according to claim 1, further comprising, after intranasally dispensing the nasal wash fluid, tilting a head of the subject farther back than while intranasally dispensing the nasal wash fluid and farther back than a neutral position.

5. The method according to claim 1, wherein intranasally dispensing the nasal wash fluid comprises intranasally dispensing the nasal wash fluid as one or more types of fluid discharge selected from the group consisting of: a fluid stream, a spray, a mist, a pressurized aerosol, and an atomized fluid.

6. The method according to claim 1, wherein intranasally dispensing the nasal wash fluid comprises intranasally dispensing the nasal wash fluid as steam.

7. The method according to claim 1, wherein intranasally dispensing the nasal wash fluid into the nasal cavity includes intranasally dispensing a total volume of 0.2-20 mL of the nasal wash fluid into the nasal cavity.

8. The method according to claim 7, wherein intranasally dispensing the nasal wash fluid into the nasal cavity comprises intranasally dispensing the total volume of the nasal wash fluid in a plurality of pulses.

9. The method according to any one of claims 1-8, wherein collecting the specimen sample comprises expressing, by the subject, the specimen sample out of the oropharynx via the anterior opening of the oral cavity.

10. The method according to claim 9, wherein expressing the specimen sample comprises spitting, by the subject, the specimen sample out of the oropharynx via the anterior opening of the oral cavity.

11. The method according to claim 9, wherein expressing the specimen sample comprises expressing the specimen sample out of the oropharynx via the anterior opening of the oral cavity after sensing, by the subject, the nasal wash fluid in the oropharynx.

12. The method according to claim 11, wherein expressing the specimen sample comprises expressing the specimen sample out of the oropharynx via the anterior opening of the oral cavity after tasting, by the subject, the nasal wash fluid in the oropharynx.

13. The method according to claim 12, wherein the nasal wash fluid includes a tastant, and wherein expressing the specimen sample comprises expressing the specimen sample out of the oropharynx via the anterior opening of the oral cavity after tasting, by the subject, the tastant in the oropharynx.

14. The method according to any one of claims 1-8, wherein collecting the specimen sample comprises drawing the specimen sample out of the oropharynx via the anterior opening of the oral cavity using an absorbent material, without swabbing the oral cavity or the oropharynx.

15. The method according to any one of claims 1-8, wherein intranasally dispensing comprises intranasally dispensing, by a healthcare worker, the nasal wash fluid into the nasal cavity.

16. The method according to any one of claims 1-8, further comprising, after intranasally dispensing the nasal wash fluid and before collecting the specimen sample, sniffing, by the subject, the biological material with the nasal wash fluid from the nasal cavity posteriorly toward the nasopharynx.

17. The method according to any one of claims 1-8, further comprising, after intranasally dispensing the nasal wash fluid and before collecting the specimen sample, snorting, by the subject, the biological material with the nasal wash fluid from the nasal cavity toward the nasopharynx.

18. The method according to any one of claims 1-8, further comprising, after collecting the specimen sample, testing the specimen sample for the presence of a particulate present in at least the biological material washed into the oropharynx by the nasal wash fluid.

19. The method according to claim 18, wherein testing the specimen sample for the presence of the particulate comprises diagnosing a disease.

20. The method according to claim 18, wherein testing the specimen sample for the presence of the particulate comprises concentrating the specimen sample by filtration prior to testing for the presence of the particulate, by passing the specimen sample through one or more filters.

21. The method according to claim 18, wherein testing the specimen sample for the presence of the particulate comprises using a molecular-based assay to test the specimen sample for the presence of the particulate.

22. The method according to claim 21, wherein testing the specimen sample for the presence of the particulate comprises using nucleic acid amplification to test the specimen sample for the presence of the particulate.

23. The method according to claim 22, wherein testing the specimen sample for the presence of the particulate comprises using polymerase chain reaction (PCR) to test the specimen sample for the presence of the particulate.

24. The method according to claim 22, wherein testing the specimen sample for the presence of the particulate comprises using isothermal amplification to test the specimen sample for the presence of the particulate.

25. The method according to claim 18, wherein testing the specimen sample for the presence of the particulate comprises performing an immunoassay to test the specimen sample for the presence of the particulate.

26. The method according to claim 18, wherein the particulate includes one or more particulates selected from the group consisting of: a pathogenic microorganism, a non-pathogenic microorganism, an antigen, a human cell, a cellular biomarker, a hormone, a chemical mediator from a cell, a pollen, a nucleic acid, and a chemical originating from an external vapor.

27. The method according to claim 18, wherein the particulate includes one or more particulates selected from the group consisting of: a virus, a bacterium, a protozoan, and a fungus.

28. The method according to claim 18, wherein the particulate includes a virus.

29. The method according to claim 28, wherein the virus is an Influenza virus.

30. The method according to claim 28, wherein the virus is a coronavirus.

31. The method according to claim 30, wherein the coronavirus is Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

32. The method according to claim 18, wherein the particulate includes a bacterium.

33. The method according to claim 32, wherein the bacterium is a Streptococcus bacterium.

34. The method according to claim 18, wherein the particulate includes one or more antigens selected from the group consisting of: a viral antigen, a bacterial antigen, and a fungal antigen.

35. The method according to any one of claims 1-8, wherein intranasally dispensing the nasal wash fluid into the nasal cavity comprises intranasally dispensing the nasal wash fluid unidirectionally within the nasal cavity.

36. The method according to any one of claims 1-8, wherein intranasally dispensing the nasal wash fluid into the nasal cavity comprises intranasally dispensing the nasal wash fluid multidirectionally within the nasal cavity.

37. The method according to claim 36, wherein intranasally dispensing the nasal wash fluid multidirectionally within the nasal cavity comprises dispensing the nasal wash fluid into the nasal cavity: in a posterior direction toward the nasopharynx, and in a superoposterior direction toward an olfactory area of the nasal cavity.

38. The method according to claim 36, wherein intranasally dispensing the nasal wash fluid multidirectionally within the nasal cavity comprises intranasally dispensing the nasal wash fluid via a plurality of nozzles.

39. The method according to claim 38, wherein intranasally dispensing the nasal wash fluid multidirectionally within the nasal cavity via the plurality of nozzles comprises intranasally dispensing the nasal wash fluid into the nasal cavity: in a posterior direction toward the nasopharynx via a first subset of one or more of the plurality of nozzles, and in a superoposterior direction toward an olfactory area of the nasal cavity via a second subset of one or more of the plurality of nozzles, the first and the second subsets not including any nozzles common to both subsets.

40. The method according to claim 39, wherein the plurality of nozzles includes only first and second nozzles, and wherein the first and the second subsets consist of the first and the second nozzles, respectively, and wherein intranasally dispensing the nasal wash fluid into the nasal cavity comprises intranasally dispensing the nasal wash fluid into the nasal cavity: in the posterior direction toward the nasopharynx via the first nozzle, and in the superoposterior direction toward the olfactory area of the nasal cavity via the second nozzle.

41. The method according to any one of claims 1-8, wherein intranasally dispensing the nasal wash fluid into the nasal cavity comprises intranasally dispensing nasal wash fluid via both nostrils of the subject simultaneously.

42. The method according to any one of claims 1-8, wherein the nasal wash fluid does not include a drug molecule.

43. The method according to any one of claims 1-8, wherein the nasal wash fluid includes a tastant.

44. The method according to any one of claims 1-8, wherein the nasal wash fluid includes a color additive, and collecting the specimen sample comprises confirming that the specimen sample contains at least a portion of the nasal wash fluid by visually ascertaining that the specimen sample includes the color additive.

45. The method according to any one of claims 1-8, further comprising, after intranasally dispensing and before collecting the specimen sample, gargling, by the subject, the nasal wash fluid washed into the oropharynx.

46. The method according to any one of claims 1-8, wherein the method further comprises, before collecting the specimen sample: orally dispensing oral wash fluid into the oral cavity via the anterior opening of the oral cavity; and swishing, by the subject, the oral wash fluid in the oral cavity, and wherein collecting comprises collecting the specimen sample that passed out of the anterior opening of the oral cavity and contains at least a portion of the oral wash fluid and the at least a portion of the biological material washed into the oropharynx with the nasal wash fluid.

47. The method according to any one of claims 1-8, wherein the method further comprises, before collecting the specimen sample: orally dispensing oral wash fluid into the oral cavity via the anterior opening of the oral cavity such that the oral wash fluid forms a mixture with the nasal wash fluid in the oropharynx; and gargling, by the subject, the mixture, and wherein collecting comprises collecting the specimen sample that passed out of the anterior opening of the oral cavity and contains at least a portion of the mixture and the at least a portion of the biological material washed into the oropharynx with the nasal wash fluid.

48. The method according to any one of claims 1-8, wherein the method further comprises, before collecting the specimen sample: orally dispensing oral wash fluid into the oral cavity via the anterior opening of the oral cavity; and gargling, by the subject, the oral wash fluid, such that the oral wash fluid forms a mixture with the nasal wash fluid, and wherein collecting comprises collecting the specimen sample that passed out of the anterior opening of the oral cavity and contains at least a portion of the mixture and the at least a portion of the biological material washed into the oropharynx with the nasal wash fluid.

49. The method according to any one of claims 1-8, wherein the method further comprises, before collecting the specimen sample: orally dispensing oral wash fluid into the oral cavity via the anterior opening of the oral cavity; and gargling, by the subject, the oral wash fluid, and wherein collecting comprises collecting the specimen sample that passed out of the anterior opening of the oral cavity and contains at least a portion of the oral wash fluid and the at least a portion of the biological material washed into the oropharynx with the nasal wash fluid.

50. The method according to any one of claims 47, 48, and 49, wherein the nasal wash fluid includes a first color additive, wherein the oral wash fluid includes a second colored additive, the first and the second color additives having different colors, and wherein collecting the specimen sample comprises confirming that the specimen sample contains at least a portion of the nasal wash fluid and at least a portion of the oral wash fluid by visually ascertaining that the specimen sample has a color produced by a combination of the first and the second color additives.

51. The method according to any one of claims 47, 48, and 49, further comprising, after collecting the specimen sample, testing the specimen sample for the presence of a particulate.

52. The method according to any one of claims 1-8, wherein the specimen sample is a first specimen sample that contains at least a first portion of the nasal wash fluid and at least a first portion of the biological material washed into the oropharynx, and wherein the method further comprises: collecting from the nasal cavity, via one or two nostrils of the subject, a second specimen sample that contains at least a second portion of the nasal wash fluid and at least a second portion of the biological material in the nasal cavity; and combining the first and the second specimen samples to produce a combined specimen sample.

53. The method according to any one of claims 1-8, wherein intranasally dispensing the nasal wash fluid into the nasal cavity comprises inserting a tubular applicator of a nasal wash fluid dispenser into a nostril of the subject and intranasally dispensing nasal wash fluid from the tubular applicator, and wherein the nasal wash fluid dispenser is shaped so as to guide introduction of the tubular applicator into the nostril at a specific orientation with respect to a maxilla of the subject.

54. The method according to any one of claims 1-8, wherein the method does not comprise swabbing the nasal cavity.

55. The method according to any one of claims 1-8, wherein collecting the specimen sample comprises drawing the specimen sample out of the oral cavity via the anterior opening of the oral cavity using an absorbent material.

56. A nasal wash fluid dispenser for intranasally dispensing nasal wash fluid into a nasal cavity of a subject, the nasal wash fluid dispenser comprising: a dispensing container, which contains the nasal wash fluid; and a tubular applicator, which (a) extends distally from and is in fluid communication with the dispensing container, (b) which is sized and shaped to be insertable into a nostril of the subject, and (c) which comprises first and second nozzles, which are configured to dispense the nasal wash fluid in first and second fluid discharges, respectively, having first and second centerlines, respectively, the first and the second centerlines defining an offset angle therebetween of between 10 and 90 degrees, such that, when the tubular applicator is inserted into the nostril with the first nozzle oriented in a posterior direction and the second nozzle oriented in a superoposterior direction, the first nozzle directs the first fluid discharge toward a nasopharynx of the subject and the second nozzle directs the second fluid discharge toward an olfactory area of the nasal cavity, such that the nasal wash fluid washes biological material into an oropharynx of the subject from (i) the nasal cavity, (ii) the nasopharynx, or (iii) the nasal cavity and the nasopharynx.

57. A nasal wash fluid dispenser for intranasally dispensing nasal wash fluid into a nasal cavity of a subject, the nasal wash fluid dispenser comprising: a dispensing container, which contains the nasal wash fluid; and a tubular applicator, which extends distally from and is in fluid communication with the dispensing container, which is sized and shaped to be insertable into a nostril of the subject, and which comprises a nozzle, which is configured to dispense the nasal wash fluid in a fluid discharge having a centerline, such that, when the tubular applicator is inserted into the nostril with the nozzle oriented in a posterior direction, the nozzle directs the fluid discharge toward a nasopharynx of the subject, so as to wash biological material, with some of the nasal wash fluid, from the nasal cavity into an oropharynx of the subject via the nasopharynx, wherein the nasal wash fluid dispenser is shaped so as to guide introduction of the tubular applicator into the nostril at a specific orientation with respect to a maxilla of the subject.

58. A nasal wash fluid system, comprising: a nasal wash fluid, which comprises a color additive; and a nasal wash fluid dispenser, which comprises: a dispensing container, which contains the nasal wash fluid; and a tubular applicator, which extends distally from and is in fluid communication with the dispensing container, and which is sized and shaped to be insertable into a nostril of a subject.

59. A method comprising: intranasally dispensing nasal wash fluid into a nasal cavity of a subject such that the nasal wash fluid washes biological material into an oropharynx of the subject from (a) the nasal cavity, (b) a nasopharynx of the subject, or (c) the nasal cavity and the nasopharynx; and thereafter, collecting a specimen sample by drawing the specimen sample out of an anterior opening of an oral cavity of the subject using an absorbent material, the specimen sample containing at least a portion of the biological material washed into the oropharynx by the nasal wash fluid.

60. A method comprising: intranasally dispensing nasal wash fluid into a nasal cavity of a subject such that the nasal wash fluid passes from the nasal cavity into an oropharynx of the subject via a nasopharynx of the subject; and thereafter, collecting a specimen sample by drawing the specimen sample out of an anterior opening of an oral cavity of the subject using an absorbent material, the specimen sample containing at least a portion of the nasal wash fluid.

61. A method comprising: intranasally dispensing nasal wash fluid into a nasal cavity of a subject such that the nasal wash fluid passes from the nasal cavity into an oropharynx of the subject via a nasopharynx of the subject; and thereafter, collecting a specimen sample that passed out of an anterior opening of an oral cavity of the subject and contains at least a portion of the nasal wash fluid.

62. A method comprising: intranasally dispensing nasal wash fluid into a nasal cavity of a subject such that the nasal wash fluid washes biological material into an oropharynx of the subject from (a) the nasal cavity, (b) a nasopharynx of the subject, or (c) the nasal cavity and the nasopharynx; and thereafter, expressing, by the subject, out of an anterior opening of an oral cavity of the subject, a specimen sample that contains at least a portion of the biological material washed into the oropharynx by the nasal wash fluid.

63. A method comprising: collecting, from a subject, a liquid specimen sample that includes saliva and an orally-dispensed oral wash fluid; passing the liquid specimen sample through a filter to trap at least some of the saliva, the filter having a nominal pore size of between 30 microns and 1.5 mm, the nominal pore size representative of a minimum size of spherical particles necessary for the filter to retain 85% of the spherical particles when H2O containing the spherical particles is passed through the filter at 20 degrees C. under pressure supplied by a 10 cm water column; and testing for the presence of a virus in the saliva trapped by the filter.

64. A method comprising: collecting, from a subject, a liquid specimen sample; passing the liquid specimen sample through a filter having a nominal pore size of between 30 microns and 1.5 mm, the nominal pore size representative of a minimum size of spherical particles necessary for the filter to retain 85% of the spherical particles when H2O containing the spherical particles is passed through the filter at 20 degrees C. under pressure supplied by a 10 cm water column; and testing for the presence of a virus that adhered to the filter during the passing of the liquid specimen sample through the filter.

65. A method comprising: collecting, from a subject, a liquid specimen sample that includes saliva and an orally-dispensed oral wash fluid; passing the liquid specimen sample through a filter to trap at least some of the saliva, the filter having a bubble-point nominal pore size of between 30 microns and 500 microns, the bubble-point nominal pore size characterized by a bubble point test using a capillary flow porometer and a wetting liquid having a fluid surface tension of 15.9 dynes/cm; and testing for the presence of a virus in the saliva trapped by the filter.

66. A method comprising: collecting, from a subject, a liquid specimen sample; passing the liquid specimen sample through a filter having a bubble-point nominal pore size of between 30 microns and 500 microns, the bubble-point nominal pore size characterized by a bubble point test using a capillary flow porometer and a wetting liquid having a fluid surface tension of 15.9 dynes/cm; and testing for the presence of a virus that adhered to the filter during the passing of the liquid specimen sample through the filter.

67. A method comprising: intranasally dispensing nasal wash fluid into a nasal cavity of a subject; thereafter, collecting a specimen sample by performing one or more nasal swabs selected from the group consisting of: an anterior nares swab and a mid-turbinate swab; and testing the specimen sample for the presence of a virus using a lateral flow immunoassay test strip.

68. A method comprising: intranasally dispensing nasal wash fluid into a nasal cavity of a subject; thereafter, tilting a head of the subject farther back than while intranasally dispensing the nasal wash fluid and farther back than a neutral position; and thereafter, collecting a specimen sample by performing one or more nasal swabs selected from the group consisting of: an anterior nares swab and a mid-turbinate swab.

69. A method comprising: intranasally dispensing nasal wash fluid into a nasal cavity of a subject; tilting a head of the subject back until a neck of the subject is fully extended; and thereafter, collecting a specimen sample by performing one or more nasal swabs selected from the group consisting of: an anterior nares swab and a mid-turbinate swab.

70. The method according to any one of claims 67, 68, and 69, wherein performing the one or more nasal swabs comprises performing the anterior nares swab.

71. The method according to claim 70, wherein intranasally dispensing the nasal wash fluid into the nasal cavity comprises: introducing, into a nostril of the subject, a swab tip of a nasal sampling device, the swab tip shaped so as to define (i) one or more nozzles, (ii) a proximal opening, and (iii) one or more channels within the swab tip that connect the one or more nozzles in fluid communication with the proximal opening; and thereafter, intranasally dispensing the nasal wash fluid into the nasal cavity from the one or more nozzles, by introducing the nasal wash fluid into the proximal opening, and wherein performing the anterior nares swab comprises performing the anterior nares swab by contacting a wall of an anterior naris of the nasal cavity with an absorbent material of the swab tip.

72. The method according to any one of claims 67, 68, and 69, wherein collecting the specimen sample comprises collecting between 25 and 150 microliters of material on a single swab.

73. The method according to claim 72, wherein collecting the specimen sample comprises collecting between 25 and 125 microliters of material on the single swab.

74. The method according to claim 72, wherein the single swab comprises a flocked swab tip, and wherein collecting the specimen sample comprises collecting between 25 and 150 microliters of the material on the flocked swab tip.

75. A method comprising: intranasally dispensing nasal wash fluid into a nasal cavity of a subject; thereafter, inserting a specimen sampler into the nasal cavity such that a farthest-inserted portion of the specimen sampler is positioned in an anterior naris of the nasal cavity; and collecting a specimen sample on the specimen sampler by rubbing a wall of the anterior naris with the specimen sampler.

76. The method according to claim 75, wherein rubbing the wall of the anterior naris comprises rotating the specimen sampler while rubbing the wall of the anterior naris with the specimen sampler.

77. The method according to claim 75, wherein the specimen sampler includes a head comprising an absorbent material, and wherein rubbing the wall of the anterior naris with the specimen sampler comprises rubbing the wall of the anterior naris with the absorbent material of the head of the specimen sampler.

78. A method comprising: intranasally dispensing nasal wash fluid into a nasal cavity of a subject; inserting a specimen sampler distally into the nasal cavity such that a farthest-inserted portion of the specimen sampler is positioned in an anterior naris of the nasal cavity; collecting a specimen sample on the specimen sampler by contacting a wall of the anterior naris with the specimen sampler; and testing the specimen sample for the presence of a virus using a lateral flow immunoassay test strip.

79. The method according to claim 78, wherein the virus is Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

80. The method according to claim 78, wherein the virus is an Influenza virus.

81. The method according to any one of claims 67-78, further comprising, after intranasally dispensing the nasal wash fluid and before collecting the specimen sample, tilting a head of the subject farther back than while intranasally dispensing the nasal wash fluid and farther back than a neutral position.

82. The method according to claim 81, further comprising, after tilting the head farther back and before collecting the specimen sample, tilting the head forward to the neutral position or a tilted-forward position.

83. The method according to claim 82, further comprising, while the subject holds the head tilted back, waiting for a period of time between (a) tilting the head farther back and (b) tilting the head forward to the neutral or the tilted-forward position, the period of time at least 2 seconds.

84. The method according to claim 83, wherein the period of time is at least 5 seconds.

85. The method according to claim 84, wherein the period of time is at least 10 seconds.

86. The method according to claim 83, wherein the period of time is a first period of time, and wherein the method further comprises, after tilting the head forward to the neutral or the tilted-forward position, waiting for a second period of time before collecting the specimen sample, the period of time at least 5 seconds.

87. The method according to claim 82, further comprising, after tilting the head forward to the neutral or the tilted-forward position, waiting for a period of time before collecting the specimen sample, the period of time at least 5 seconds.

88. The method according to claim 87, wherein the period of time is at least 10 seconds.

89. The method according to claim 82, wherein tilting the head forward comprises tilting the head forward to the neutral or the tilted-forward position after sensing, by the subject, the nasal wash fluid in an oropharynx of the subject.

90. The method according to any one of claims 67-78, wherein intranasally dispensing the nasal wash fluid comprises: tilting a head of the subject back until a neck of the subject is fully extended; and while the subject holds the head tilted back with the neck fully extended, intranasally dispensing nasal wash fluid into a nasal cavity of the subject.

91. The method according to claim 90, further comprising, after intranasally dispensing the nasal wash, tilting the head forward to a neutral position or a tilted-forward position.

92. The method according to claim 91, further comprising, while the subject holds the head tilted back with the neck fully extended, waiting for a period of time between (a) concluding intranasally dispensing the nasal wash fluid into the nasal cavity and (b) tilting the head forward to the neutral or the tilted-forward position, the period of time at least 2 seconds.

93. The method according to claim 92, wherein the period of time is at least 5 seconds.

94. The method according to claim 93, wherein the period of time is at least 10 seconds.

95. The method according to claim 92, wherein the period of time is a first period of time, and wherein the method further comprises, after tilting the head forward to the neutral or the tilted-forward position, waiting for a second period of time before collecting the specimen sample, the period of time at least 5 seconds.

96. The method according to claim 91, further comprising, after tilting the head forward to the neutral or the tilted-forward position, waiting for a period of time before collecting the specimen sample, the period of time at least 5 seconds.

97. The method according to claim 96, wherein the period of time is at least 10 seconds.

98. The method according to any one of claims 67-78, wherein intranasally dispensing the nasal wash fluid comprises intranasally dispensing the nasal wash fluid into the nasal cavity while a head of the subject is held in a neutral position or a tilted-forward position.

99. The method according to any one of claims 67-78, further comprising, after intranasally dispensing the nasal wash fluid and before collecting the specimen sample, sniffing, by the subject, the nasal wash fluid.

100. A method comprising: intranasally dispensing nasal wash fluid into a nasal cavity of a subject; thereafter, tilting a head of the subject farther back than while intranasally dispensing the nasal wash fluid and farther back than a neutral position; inserting a specimen sampler into the nasal cavity such that a farthest-inserted portion of the specimen sampler is positioned in an anterior naris of the nasal cavity; and collecting a specimen sample on the specimen sampler by contacting a wall of the anterior naris with the specimen sampler.

101. The method according to claim 100, wherein tilting the head farther back comprises tilting the head back until a neck of the subject is fully extended.

102. The method according to claim 100, further comprising, after tilting the head farther back and before collecting the specimen sample, tilting the head forward to the neutral position or a tilted-forward position.

103. The method according to claim 102, further comprising, while the subject holds the head tilted back, waiting for a period of time between (a) tilting the head farther back and (b) tilting the head forward to the neutral or the tilted-forward position, the period of time at least 2 seconds.

104. The method according to claim 103, wherein the period of time is at least 5 seconds.

105. The method according to claim 104, wherein the period of time is at least 10 seconds.

106. The method according to claim 103, wherein the period of time is a first period of time, and wherein the method further comprises, after tilting the head forward to the neutral or the tilted-forward position, waiting for a second period of time before collecting the specimen sample, the period of time at least 5 seconds.

107. The method according to claim 102, further comprising, after tilting the head forward to the neutral or the tilted-forward position, waiting for a period of time before collecting the specimen sample, the period of time at least 5 seconds.

108. The method according to claim 107, wherein the period of time is at least 10 seconds.

109. The method according to claim 102, wherein tilting the head forward comprises tilting the head forward to the neutral or the tilted-forward position after sensing, by the subject, the nasal wash fluid in an oropharynx of the subject.

110. The method according to claim 100, wherein intranasally dispensing the nasal wash fluid comprises intranasally dispensing the nasal wash fluid into the nasal cavity while the head is held in the neutral position or a tilted-forward position.

111. The method according to claim 100, further comprising, after intranasally dispensing the nasal wash fluid and before collecting the specimen sample, sniffing, by the subject, the nasal wash fluid.

112. A method comprising: tilting a head of a subject back until a neck of the subject is fully extended; while the subject holds the head tilted back with the neck fully extended, intranasally dispensing nasal wash fluid into a nasal cavity of the subject; inserting a specimen sampler into the nasal cavity such that a farthest-inserted portion of the specimen sampler is positioned in an anterior naris of the nasal cavity; and collecting a specimen sample on the specimen sampler by contacting a wall of the anterior naris with the specimen sampler.

113. The method according to claim 112, further comprising, after intranasally dispensing the nasal wash, tilting the head forward to a neutral position or a tilted-forward position.

114. The method according to claim 113, further comprising, while the subject holds the head tilted back with the neck fully extended, waiting for a period of time between (a) concluding intranasally dispensing the nasal wash fluid into the nasal cavity and (b) tilting the head forward to the neutral or the tilted-forward position, the period of time at least 2 seconds.

115. The method according to claim 114, wherein the period of time is at least 5 seconds.

116. The method according to claim 115, wherein the period of time is at least 10 seconds.

117. The method according to claim 114, wherein the period of time is a first period of time, and wherein the method further comprises, after tilting the head forward to the neutral or the tilted-forward position, waiting for a second period of time before collecting the specimen sample, the period of time at least 5 seconds.

118. The method according to claim 113, further comprising, after tilting the head forward to the neutral or the tilted-forward position, waiting for a period of time before collecting the specimen sample, the period of time at least 5 seconds.

119. The method according to claim 118, wherein the period of time is at least 10 seconds.

120. The method according to claim 112, further comprising, after intranasally dispensing the nasal wash fluid and before collecting the specimen sample, sniffing, by the subject, the nasal wash fluid.

121. The method according to any one of claims 78-120, wherein the specimen sampler includes a head comprising an absorbent material, and wherein contacting the wall of the anterior naris with the specimen sampler comprises contacting the wall of the anterior naris with the absorbent material of the head of the specimen sampler.

122. The method according to any one of claims 75-120, wherein the specimen sampler comprises a swab.

123. The method according to claim 122, wherein the swab comprises a flocked swab tip.

124. The method according to claim 122, wherein the swab comprises a foam swab tip.

125. The method according to claim 122, wherein the swab comprises a tip having a greatest diameter of between 2 and 5 mm.

126. The method according to claim 122, wherein the swab comprises a tip having a collection surface area of between 200 and 300 mm2.

127. The method according to claim 122, wherein the swab comprises a tip having a volume of between 200 and 300 mm3.

128. The method according to claim 122, wherein the swab comprises a tip having a length of between 12 and 20 mm.

129. The method according to any one of claims 75-120, wherein collecting the specimen sample comprises collecting the specimen sample on the specimen sampler without using suction.

130. The method according to any one of claims 67-120, wherein intranasally dispensing the nasal wash fluid comprises intranasally dispensing the nasal wash fluid without blowing, by the subject, a nose of the subject immediately prior to intranasally dispensing the nasal wash fluid.

131. The method according to any one of claims 75-120, wherein collecting the specimen sample on the specimen sampler comprises collecting between 25 and 150 microliters of material on the specimen sampler.

132. The method according to claim 131, wherein collecting the specimen sample on the specimen sampler comprises collecting between 25 and 125 microliters of material on the specimen sampler.

133. The method according to claim 131, wherein the specimen sampler comprises a swab comprising a flocked swab tip, and wherein collecting the specimen sample on the specimen sampler comprises collecting between 25 and 150 microliters of the material on the flocked swab tip.

134. The method according to any one of claims 67-120, further comprising, after intranasally dispensing the nasal wash fluid into the nasal cavity: orally dispensing oral wash fluid into an oral cavity of the subject via an anterior opening of the oral cavity; and gargling, by the subject, the oral wash fluid.

135. The method according to any one of claims 67-120, wherein intranasally dispensing the nasal wash fluid comprises intranasally dispensing the nasal wash fluid as one or more types of fluid discharge selected from the group consisting of: a spray, a mist, a pressurized aerosol, steam, and an atomized fluid.

136. The method according to any one of claims 67-120, wherein intranasally dispensing the nasal wash fluid comprises intranasally dispensing the nasal wash fluid as a fluid stream.

137. The method according to any one of claims 67-120, wherein collecting the specimen sample comprises collecting the specimen sample after sensing, by the subject, the nasal wash fluid in an oropharynx of the subject.

138. The method according to any one of claims 68-77 and 801-821, further comprising testing the specimen sample for the presence of a particulate.

139. The method according to claim 138, wherein the particulate is selected from the group consisting of: a virus, a bacterium, a protozoan, and a fungus.

140. The method according to claim 139, wherein the particulate is the virus.

141. The method according to claim 140, wherein the virus is Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

142. The method according to claim 140, wherein the virus is an Influenza virus.

143. The method according to claim 140, wherein testing for the presence of the virus comprises using nucleic acid amplification.

144. The method according to claim 143, wherein testing for the presence of the virus comprises using polymerase chain reaction (PCR).

145. The method according to claim 140, wherein testing for the presence of the virus comprises performing an immunoassay.

146. The method according to claim 145, wherein performing the immunoassay comprises using a lateral flow immunoassay test strip.

147. The method according to claim 146, wherein performing the immunoassay comprises performing a chromatographic digital immunoassay.

148. A method comprising: intranasally dispensing nasal wash fluid into a nasal cavity of a subject; and collecting a specimen sample by performing one or more nasal swabs selected from the group consisting of: an anterior nares swab, a mid-turbinate swab, and a nasopharyngeal swab.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0404] FIG. 1 is a schematic illustration of a nasal wash fluid dispenser, in accordance with an application of the present invention;

[0405] FIGS. 2A-D are schematic cross-sectional illustrations of a method of collecting a specimen sample from a subject, in accordance with an application of the present invention;

[0406] FIGS. 3A-B are schematic cross-sectional illustrations of the nasal wash fluid dispenser of FIG. 1 inserted into a nasal cavity, in accordance with an application of the present invention;

[0407] FIG. 4 is a schematic cross-sectional illustration of a portion of a method for collecting a specimen sample from the subject, in accordance with an application of the present invention;

[0408] FIGS. 5A-B are schematic illustrations of a portion of a method for collecting a specimen sample from a subject, in accordance with an application of the present invention;

[0409] FIG. 6 is a schematic illustration of the nasal wash fluid dispenser of FIG. 1 with a tubular applicator thereof inserted into a nostril, in accordance with an application of the present invention;

[0410] FIGS. 7A-K are several schematic illustrations of the nasal wash fluid dispenser of FIG. 1 viewed from several different directions, in accordance with an application of the present invention;

[0411] FIGS. 8A-C are schematic illustrations of another nasal wash fluid dispenser, in accordance with an application of the present invention;

[0412] FIGS. 9A-F are schematic illustrations of yet another nasal wash fluid dispenser, in accordance with an application of the present invention;

[0413] FIG. 10 is a schematic cross-sectional illustration of the nasal wash fluid dispenser of FIGS. 9A-F inserted into the nasal cavity, in accordance with an application of the present invention;

[0414] FIG. 11 is a schematic cross-sectional illustration of a tubular applicator of the nasal wash fluid dispenser of FIGS. 9A-F, in accordance with an application of the present invention;

[0415] FIG. 12 is a schematic illustration of a testing kit, in accordance with an application of the present invention;

[0416] FIG. 13 is a schematic illustration of another testing kit, in accordance with an application of the present invention;

[0417] FIGS. 14A-B are schematic illustrations of filter units for testing a liquid specimen sample that includes saliva for the presence of a particulate, in accordance with respective applications of the present invention;

[0418] FIG. 15 is a schematic illustration of a testing kit, in accordance with an application of the present invention;

[0419] FIG. 16 is a schematic illustration of another testing kit, in accordance with an application of the present invention;

[0420] FIGS. 17A-B are schematic illustrations of a method for collecting a specimen sample from a subject, in accordance with an application of the present invention;

[0421] FIG. 18 is a schematic illustration of a testing kit, in accordance with an application of the present invention;

[0422] FIG. 19 is a schematic illustration of another testing kit, in accordance with an application of the present invention;

[0423] FIGS. 20A-B and 21A-D are schematic illustrations of respective configurations of a nasal sampling device for collecting a specimen sample, in accordance with respective applications of the present invention;

[0424] FIG. 22 is a schematic illustration of the use of nasal sampling device of FIGS. 20A-B and 21A-B for collecting a specimen sample from a nasal cavity, in accordance with an application of the present invention;

[0425] FIGS. 23A and 23B are schematic illustrations of kits that comprise portions of the nasal sampling devices of FIGS. 20A-B and 21A-B, respectively, and a vial, in accordance with respective applications of the present invention;

[0426] FIGS. 24A and 24B are schematic illustrations of another testing kit, in accordance with an application of the present invention;

[0427] FIGS. 25A-B are charts and FIG. 25C is a table that present results of an experiment conducted on behalf of the inventors, described hereinbelow as Experiment 8, in accordance with an application of the present invention;

[0428] FIG. 26A is a chart and FIG. 26B is a table that present results of the experiment conducted on behalf of the inventors, described hereinbelow as Experiment 8, in accordance with an application of the present invention; and

[0429] FIGS. 27A-B are charts that present results of the experiment conducted on behalf of the inventors, described hereinbelow as Experiment 8, in accordance with an application of the present invention.

DETAILED DESCRIPTION OF APPLICATIONS

[0430] Reference is made to FIG. 1, which is a schematic illustration of a nasal wash fluid dispenser 20, in accordance with an application of the present invention.

[0431] Reference is also made to FIGS. 2A-D, which are schematic cross-sectional illustrations of a method of collecting a specimen sample 24 from a subject, for example one or more specimen samples 24, in accordance with an application of the present invention.

[0432] As shown in FIGS. 2A-B, nasal wash fluid 26 is intranasally dispensed into a nasal cavity 22 of the subject, via one or both nostrils 28. In order to intranasally dispense nasal wash fluid 26, a nasal dispenser is inserted into one or both nostrils 28. For example, the nasal dispenser may comprise a conventional nasal dispenser known in the art (also known as a nasal applicator), a conventional nasal dropper known in the art, or a conventional ampoule known in the art, all of which are inserted into one nostril at a time. Alternatively, the nasal dispenser may comprise one of the nasal dispensers described herein, such as nasal wash fluid dispenser 20 or the other nasal dispensers described hereinbelow, which may be inserted into one or both nostrils 28. FIG. 2A shows nasal wash fluid dispenser 20 inserted into nasal cavity 22 via one of nostrils 28.

[0433] As shown in FIG. 2C, nasal wash fluid 26 is intranasally dispensed such that nasal wash fluid 26 washes biological material (typically, but necessarily, including saliva, which contains inter alia mucus) into an oropharynx 30 from (a) nasal cavity 22, (b) a nasopharynx 32, or (c) nasal cavity 22 and nasopharynx 32. Typically, nasal wash fluid 26 loosens the biological material in nasal cavity 22, nasopharynx 32, and/or oropharynx 30.

[0434] As shown in FIG. 2D, the method further comprises, thereafter, collecting, via an anterior opening 34 of an oral cavity 36 of the subject, specimen sample 24 that contains at least a portion of the biological material washed into oropharynx 30 by nasal wash fluid 26. (Anterior opening 34 is the opening of the mouth between the lips, between outside oral cavity 36 and inside oral cavity 36.)

[0435] Specimen sample 24 is typically collected into a collection receptacle 40, as shown in FIG. 2D.

[0436] For some applications, collection receptacle 40 may have a surface that is treated so as not to allow sticking of a particulate.

[0437] In some applications, collection receptacle 40 used to collect specimen sample 24 comprises a stabilizer (e.g., to help maintain enzyme activity), an inhibitor (e.g., to help prevent RNA degradation), and/or a purifier. Alternatively or additionally, in some applications, after collection, specimen sample 24 is transferred to a liquid container of a testing device, such as any of the testing devices described in the patent applications incorporated herein by reference below; the liquid container of the testing device comprises a stabilizer, an inhibitor, and/or a purifier.

[0438] This method comprehensively samples multiple respiratory sites of the upper respiratory tract to collect biological material in a specimen sample 24 and optionally can be performed by the subject (patient) alone without exposing others to contagion. Alternatively, the method can be performed entirely or partially by somebody other than the subject, for example a healthcare provider or a relative of the subject. The method is simpler to perform than nasopharyngeal and pharyngeal swabbing conventionally performed for sample specimen collection, which are heavily dependent on the skill of the healthcare provider and are believed to result in many false negatives because of poor techniques. In addition, conventional swabbing cannot be performed by most subjects on themselves.

[0439] In some applications, the method comprises collection of one or more specimen samples 24 containing biological material from one or more of the following locations: nasal cavity 22, nasopharynx 32, and oropharynx 30, and, in some applications (such as when the method further comprises gargling and/or swishing, as described hereinbelow), a posterior pharynx 42, a hypopharynx 44 and lower airways, and in some applications, oral cavity 36, thereby sampling the entire area where pathogens are found in the upper aerodigestive tract.

[0440] In applications in which the particulate includes a virus, the virus may include free virions (i.e., extracellular virions) and/or intracellular virions, which, optionally are released from cells (e.g., epithelial cells) of the biological material after collection of specimen sample 24 and before testing the presence of the virus. For example, testing techniques may be used that are described in U.S. Provisional Application 62/988,259, filed Mar. 11, 2020, and/or International Application PCT/IB2021/052055, filed Mar. 11, 2021, which are assigned to the assignee of the present application and incorporated herein by reference.

[0441] Reference is still made to FIGS. 2A-D. As used in the present application, including in the claims and Inventive Concepts, “biological material” includes, but is not limited to, microorganisms, antigens, human cells, extracellular soluble and non-soluble particles, cellular components and products, blood products, cellular biomarkers, hormones, vitamins, electrolytes, chemical mediators from cells such as mediators of inflammation, pollens, mucus, saliva, sputum, respiratory particles, droplets derived from the upper and lower airways, nucleic acids including DNA and RNA, and chemicals originating from external vapors. As used in the present application, including in the claims and Inventive Concepts, microorganisms may include either pathogenic microorganisms or non-pathogenic microorganisms or both, for example, viruses, bacteria, protozoa, and fungi.

[0442] As used in the present application, including in the claims and Inventive Concepts, human cells may include epithelial cells, for example, goblet cells and/or columnar epithelial cells primarily derived from nasal cavity 22 and/or squamous epithelial cells primarily derived from oral cavity 36. As used in the present application, including in the claims and Inventive Concepts, human cells may also include cellular responders of the immune system, for example, leukocytes (e.g., neutrophils, eosinophils, lymphocytes, or monocytes), macrophages, mast cells, and histocytes. As used in the present application, including in the claims and Inventive Concepts, cellular components and products include lipids, proteins, glycoproteins, proteoglycans, mucins, and nucleic acids such as DNA and RNA.

[0443] Reference is again made to FIG. 2C. For some applications, the subject tilts his or her head back before, while, and/or after nasal wash fluid 26 is intranasally dispensed into nasal cavity 22, e.g., such as described in more detail hereinbelow with reference to FIGS. 17A-B. Alternatively, the subject holds his or her head in a neutral position or tilted forward while nasal wash fluid 26 is intranasally dispensed into nasal cavity 22, and optionally tilts his or her head back after nasal wash fluid 26 is intranasally dispensed into nasal cavity 22. Although tilting of the subject's head back and forward is generally described herein as being performed by the subject himself or herself, the subject's head may alternatively be tilted by somebody else, such as if the subject is too young or otherwise unable to tilt his or her own head.

[0444] For any of the applications described herein, intranasally dispensing nasal wash fluid 26 into nasal cavity 22 may comprise intranasally dispensing nasal wash fluid 26 into nasal cavity 22 while the subject is lying supine. For example, this position may be particularly appropriate when a conventional nasal dropper or a conventional ampoule is used for dispending nasal wash fluid 26.

[0445] In some applications, the method further comprises waiting for a period of time between (a) concluding intranasally dispensing nasal wash fluid 26 into nasal cavity 22 and (b) collecting specimen sample 24, the period of time at least 1 minute, e.g., at least 2 minutes, at least 3 minutes, or at least 5 minutes, such as at least 10 minutes. Optionally, the period of time is at least 15 minutes, or at least 30 minutes. The inventors found that waiting for a period of time less than 90 minutes between intranasally dispensing nasal wash fluid 26 into nasal cavity 22 and collecting specimen sample 24 increased the quantity of virus in the collected specimen sample. The inventors hypothesize that waiting for the period of time provides enough time for the subject's natural nasal mucociliary clearance (NMC) system to transport nasal wash fluid 26 containing biological material (typically including mucus) from nasal cavity 22 into oropharynx 30.

[0446] Reference is again made to FIG. 2B. In some applications, the method comprises dispensing intranasally a total volume of 0.2-20 mL of a nasal wash fluid 26, for example, 0.2-10 mL, for example, 0.2-5 mL, for example, 0.6-5 mL, into nasal cavity 22. The total nasal wash fluid 26 volume of 0.2-20 mL describes the total volume introduced into the entire nasal cavity 22 through either one or both nostrils 28. For example, 0.1-10 mL may be dispensed into each nostril 28, for a total volume of 0.2-20 mL, or for example, 0.2-20 mL may be dispensed into one nostril 28. Experiments performed by the inventors showed that fluid drained into oropharynx 30 when 1 mL of fluid was dispensed into nasal cavity 22, and did not drain into oropharynx 30 enough to be expressed out of the mouth when only 0.5 mL of fluid was dispensed into nasal cavity 22. However, the inventors hypothesize that dispensing a smaller volume of nasal wash fluid 26, such as as little as 0.2 mL, into nasal cavity 22 may be adequate for draining into oropharynx 30 and expressing out of the mouth if nasal wash fluid 26 is dispensed at a higher pressure and/or is better aimed toward nasopharynx 32 than in the experiments performed.

[0447] In some applications, nasal wash fluid 26 is intranasally dispensed as one or more types of fluid discharge selected from the group consisting of: a fluid stream, a spray, a mist, a pressurized aerosol, steam, and an atomized fluid.

[0448] In some applications, nasal wash fluid 26 is dispensed intranasally using a circular rotary motion that generates dispersion within nasal cavity 22.

[0449] In some applications, nasal wash fluid 26 is intranasally dispensed in a plurality of pulses so as to increase volume without sacrificing comfort; for example, each of the pulses may have a volume of 0.02-10 mL, for example, 0.02-1 mL. Lowering the volume or the pressure of the fluid pulses reduces discomfort and unintentional swallowing, and a plurality of pulses increases volume and surface area sampled, thereby increasing the amount of total particulate collected in specimen sample 24, for improved sampling.

[0450] In some applications, nasal wash fluid 26 comprises a non-irritant solution; for example, the non-irritant solution may comprise or consist of water. In some applications, the non-irritant solution comprises saline solution that may be hypertonic, isotonic, or hypotonic, for example, a phosphate-buffered saline solution.

[0451] Typically, nasal wash fluid 26 does not comprise a drug molecule. Alternatively, nasal wash fluid 26 does comprise a drug molecule.

[0452] For some applications, nasal wash fluid 26 comprises an odorant.

[0453] Reference is made to FIGS. 2B and 2C. In some applications, nasal wash fluid 26 is intranasally dispensed into nasal cavity 22 (either continuously or in a plurality of pulses) until the subject senses (e.g., by feeling and/or tasting) nasal wash fluid 26 in oropharynx 30. For applications which comprise gargling and/or swishing, as described herein, the subject typically gargles and/or swishes after sensing nasal wash fluid 26 in oropharynx 30.

[0454] For some of these applications, nasal wash fluid 26 comprises a tastant, for example, menthol flavoring, to monitor fluid passage and the movement of the biological material and nasal wash fluid 26 from nasal cavity 22 and nasopharynx 32 into oropharynx 30. Optionally, as mentioned above, the subject tilts his or her head back before, while, and/or after nasal wash fluid 26 is intranasally dispensed into nasal cavity 22, e.g., such as described in more detail hereinbelow with reference to FIGS. 17A-B. Further optionally, for applications in which nasal wash fluid 26 is separately intranasally dispensed into both nostrils, after the subject senses, in oropharynx 30, nasal wash fluid 26 administered to one of the nostrils, nasal wash fluid is intranasally dispensed into the other nostril, until the subject senses the additional nasal wash fluid in oropharynx 30.

[0455] As used in the present application, including in the claims and Inventive Concepts, a “tastant” is a taste-provoking chemical molecule that is dissolved in a fluid, and stimulates the sense of taste. For example, the tastant may comprise a non-salty flavor, such as menthol flavoring, a sweet flavor (e.g., comprising a sugar), a bitter flavor; a salty flavor, e.g., saline solution; and/or a non-bitter flavor.

[0456] Alternatively or additionally, in some applications, nasal wash fluid 26 comprises a color additive to monitor fluid passage and the movement of the biological material and nasal wash fluid 26 from nasal cavity 22 and nasopharynx 32 into oropharynx 30, and collecting specimen sample 24 comprises confirming that specimen sample 24 contains at least a portion of nasal wash fluid 26 by visually ascertaining that specimen sample 24 comprises the color additive. For some applications, visually ascertaining is performed by the human eye. Alternatively, for some applications, visually ascertaining is performed using spectroscopy, image processing, and/or three-dimensional (3D) image sensing; any of these techniques may optionally utilize artificial intelligence. Such confirmation may identify an inadvertent and/or a deliberate lack of administration of the nasal wash and/or passage of the nasal wash into oropharynx 30.

[0457] Alternatively or additionally, in some applications, nasal wash fluid 26 comprises a chemical additive (which may or may not have a color) to monitor fluid passage and the movement of the biological material and nasal wash fluid 26 from nasal cavity 22 and nasopharynx 32 into oropharynx 30, and collecting specimen sample 24 comprises confirming that specimen sample 24 contains at least a portion of nasal wash fluid 26 by ascertaining that specimen sample 24 includes the chemical additive. For some applications, ascertaining is performed using spectroscopy, image processing, and/or three-dimensional (3D) image sensing; any of these techniques may optionally utilize artificial intelligence. For some applications, ascertaining is performed using the same technique used to test for the presence of the particulate, and the chemical additive is configured to be detectable during the test (for example, the chemical additive may be a marker, optionally including DNA and/or RNA). For example, ascertaining may be performed using an immunoassay (for example, a lateral-flow immunochromatographic assay, a chromatographic digital immunoassay, or an enzyme-linked immunosorbent assay (ELISA)); or a molecular-based assay (for example, nucleic acid hybridization, or for example, nucleic acid amplification, including polymerase chain reaction (PCR) amplification, real-time quantitative PCR (qPCR) amplification, reverse transcriptase PCR (RT-PCR) amplification, and isothermal amplification, such as loop-mediated isothermal amplification (LAMP)). Such confirmation may identify an inadvertent and/or a deliberate lack of administration of the nasal wash and/or passage of the nasal wash into oropharynx 30.

[0458] Further alternatively or additionally, in some applications, nasal wash fluid 26 comprises a chemical that increases the amount of biological material in specimen sample 24 by enhancing the clearance of the mucus as well as dislodging the biological material lining the mucosa, for example, a mucolytic agent such as acetylcysteine, or for example, a detergent such as polysorbate, or for example, a gas such as pressurized carbon dioxide to generate bubbles or nitrous oxide to increases the speed of mucociliary clearance.

[0459] Reference is made to FIGS. 3A-B, which are schematic cross-sectional illustrations of nasal wash fluid dispenser 20 inserted into nasal cavity 22, in accordance with an application of the present invention. FIG. 3B shows nasal wash fluid dispenser 20 rotated slightly along a lateral axis compared to FIG. 3A, so as to provide an additional perspective.

[0460] In some applications, the method described hereinabove with reference to FIGS. 2A-D comprises dispensing nasal wash fluid 26 multidirectionally into each of the one or two fossae of nasal cavity 22. In some applications, intranasally dispensing nasal wash fluid 26 multidirectionally within nasal cavity 22 comprises dispensing nasal wash fluid 26 into nasal cavity 22 in a posterior direction 48 toward nasopharynx 32 and in a superoposterior direction 50 toward an olfactory area 52 of nasal cavity 22 above a superior nasal concha 54, for example, through a plurality of nozzles 60, for example, through exactly two nozzles 60.

[0461] As used in the present application, including in the claims and Inventive Concepts, a “nozzle” is a device designed to control the direction or characteristics of a fluid flow as it exits an enclosed chamber or tube.

[0462] In some applications, intranasally dispensing nasal wash fluid 26 multidirectionally within nasal cavity 22 comprises intranasally dispensing nasal wash fluid 26 within nasal cavity 22 via the plurality of nozzles 60 with respective different fluid flow characteristics, the fluid flow characteristics including one or more characteristics selected from the group consisting of: volume, sprayed particle size, velocity, and flow rate. For example, in some applications, the volume of nasal wash fluid 26 dispensed in posterior direction 48 may be greater than the volume of nasal wash fluid 26 dispensed in superoposterior direction 50 so as to increase the amount of biological material collected in specimen sample 24, since the majority of the biological material contained within nasal cavity 22 drains towards nasopharynx 32 and therefore is believed to be a prime site for sampling. In other applications, for example, the volume of nasal wash fluid 26 dispensed in superoposterior direction 50 may be greater than the volume of nasal wash fluid 26 dispensed in posterior direction 48. These applications may be beneficial since the inventors hypothesize that some viruses may be more concentrated in olfactory area 52, and that reflexive swallowing or premature spitting of nasal wash fluid 26 by the subject may sometimes be induced if a large volume of fluid is dispensed posteriorly.

[0463] In some applications, intranasally dispensing nasal wash fluid 26 multidirectionally within nasal cavity 22 comprises intranasally dispensing nasal wash fluid 26 within nasal cavity 22 via the plurality of nozzles 60 with equal fluid flow characteristics, the fluid flow characteristics including one or more characteristics selected from the group consisting of: volume, sprayed particle size, velocity, and flow rate.

[0464] In some applications, intranasally dispensing nasal wash fluid 26 multidirectionally within nasal cavity 22 comprises intranasally dispensing nasal wash fluid 26 within nasal cavity 22 via the plurality of nozzles 60 either simultaneously or sequentially.

[0465] In some applications, the method described hereinabove with reference to FIGS. 2A-D comprises dispensing nasal wash fluid 26 unidirectionally into both nostrils 28 (and thus into both fossae of the nasal cavity), for example, through a single nozzle 60. In some applications, intranasally dispensing nasal wash fluid 26 unidirectionally within nasal cavity 22 comprises dispensing nasal wash fluid 26 into nasal cavity 22 in posterior direction 48 toward nasopharynx 32 near a pharyngeal recess 58, or in superoposterior direction 50 toward olfactory area 52, or in a superior direction 62. For these applications, nasal wash fluid dispenser 20 may comprise only a single nozzle 60 (configuration not shown), or a conventional nasal dispenser known in the art may be used.

[0466] Dispensing nasal wash fluid 26 to target the posterior aspect of nasal cavity 22 toward nasopharynx 32 may increase the quality of the collected specimen sample 24 because a majority of the biological material contained within nasal cavity 22 drains toward nasopharynx 32, which is therefore a prime site for sampling. Alternatively or additionally, dispensing nasal wash fluid 26 to target the superoposterior aspect of nasal cavity 22 toward olfactory area 52 may increase the quality of the collected specimen sample 24 because olfactory area 52 contains high levels of infectious particles in infected individuals, such as with viruses, for example, SARS-CoV-2, in infected individuals with or without symptoms of olfactory dysfunction. A case study analyzed MRIs of a patient with confirmed COVID-19 who had experienced symptomatic olfactory dysfunction and determined that there were no anomalies present in the olfactory bulb and track, but that there was significant bilateral inflammation of the olfactory cleft in the nasal cavity. (Eliezer M, Hautefort C, Hamel A, et al. (2020). “Sudden and Complete Olfactory Loss Function as a Possible Symptom of COVID-19.” JAMA Otolaryngol Head Neck Surg. Published online Apr. 8, 2020. doi:10.1001/jamaoto.2020.0832). This case study supports the hypothesis that olfactory area 52 around the olfactory cleft above superior nasal concha 54 in nasal cavity 22 may contain high levels of infectious particles in individuals infected with SARS-CoV-2.

[0467] In some applications, intranasally dispensing nasal wash fluid into nasal cavity 22 comprises inserting a tubular applicator 64 of nasal wash fluid dispenser 20 into a nostril 28 of the subject, for example, inserting two tubular applicators 64 of nasal wash fluid dispenser 20 into both nostrils 28 of the subject.

[0468] Reference is again made to FIGS. 2B-C. In some applications, after nasal wash fluid 26 is dispensed into nasal cavity 22, the subject then sniffs or snorts, one or more times, the intranasally-introduced nasal wash fluid 26 posteriorly further into nasal cavity 22, allowing nasal wash fluid 26 to reach nasopharynx 32, from where nasal wash fluid 26 then drains into oropharynx 30. In other words, the sniffing or snorting moves nasal wash fluid 26 within nasal cavity 22 posteriorly toward nasopharynx 32 from where it subsequently drains to oropharynx 30. The sniffing or snorting comprises swiftly drawing air through the nose in short, usually audible, inhalations, that are more sudden and stronger than ordinary breathing. The sniffing or snorting does not necessarily comprise smelling. Alternatively, the intranasally-introduced nasal wash fluid 26 moves posteriorly further into nasal cavity 22 to reach nasopharynx 32 through normal mucociliary movement, without any special action by the subject, such as by sniffing or snorting. Nasal wash fluid 26 then drains from nasopharynx 32 into oropharynx 30, such as shown in FIG. 2C. In other words, the normal mucociliary movement moves nasal wash fluid 26 within nasal cavity 22 posteriorly toward nasopharynx 32, from where nasal wash fluid 26 subsequently drains to oropharynx 30.

[0469] In some applications, collecting specimen sample 24 from oral cavity 36 of the subject is performed by the subject alone, while in other applications, collecting specimen sample 24 from oral cavity 36 of the subject is performed by someone other than the subject, for example, by a healthcare provider or relative of the subject.

[0470] Reference is again made to FIG. 2D. In some applications, collecting specimen sample 24 comprises collecting specimen sample 24 from oral cavity 36 of the subject by the subject expressing, e.g., spitting, out the contents of oral cavity 36 from oral cavity 36 via anterior opening 34. Typically, the contents contain at least a portion of the intranasally-introduced nasal wash fluid 26 and at least a portion of the biological material. For example, the subject may spit out at least a portion of the intranasally-introduced nasal wash fluid 26 and at least a portion of an orally-introduced oral wash fluid 68 from oral cavity 36. In some applications, collecting the one or more specimen samples 24 comprises collecting a specimen sample 24 that was spit, by the subject, out of oral cavity 36 via anterior opening 34, which typically contains at least a portion of the intranasally-introduced nasal wash fluid 26 and at least a portion of the biological material.

[0471] Typically, in applications in which specimen sample 24 is collected by the subject expressing, e.g., spitting, out the contents of oral cavity 36, specimen sample 24 is collected into collection receptacle 40, such as by the subject directly expressing, e.g., spitting, specimen sample 24 into collection receptacle 40. Collection receptacle 40 may, for example, comprise a cup, as shown, or may alternatively comprise a collection receptacle of a sample processing device or sampling testing device, such as described, for example, in one or more of the patent applications incorporated hereinbelow by reference, or otherwise known in the art.

[0472] For any of the applications described herein that comprise spitting by the subject, the spitting optionally comprises hawking, i.e., forcefully spitting after forcing material up from the throat, e.g., by making a harsh guttural sound.

[0473] Reference is now made to FIG. 4, which is a schematic cross-sectional illustration of a portion of a method for collecting specimen sample 24 from the subject, in accordance with an application of the present invention. In some applications, the method of FIGS. 2A-D further comprises, before collecting specimen sample 24, spraying, via anterior opening 34 of oral cavity 36, oropharynx 30 with oropharyngeal fluid wash 46. This spraying may help loosen and increase the collection of biological material from the walls of oropharynx 30 (e.g., the walls of posterior pharynx 42), and/or may provide better coverage of posterior pharynx 42 than gargling and/or swishing alone, and thus result in better sampling of posterior pharynx 42 than gargling and/or swishing alone. This spraying is typically performed after intranasally dispensing nasal wash fluid 26 into nasal cavity 22, but may alternatively be performed prior to intranasally dispensing nasal wash fluid 26 into nasal cavity 22.

[0474] Reference is again made to FIGS. 2A-D. In some applications, the nasally-introduced nasal wash fluid 26, after it has been washed into oropharynx 30, such as shown in FIG. 2C, is gargled by the subject, swished by the subject in oral cavity 36, or both gargled and swished by the subject in oral cavity 36, prior to collecting specimen sample 24 from oral cavity 36, such as shown in FIG. 2D. In some of these applications, the subject alternates multiple times between (a) gargling and/or swishing and (b) sniffing or snorting, before collecting specimen sample 24 from oral cavity 36. Typically, the alternation between (a) gargling and/or swishing and (b) sniffing or snorting occurs while nasal wash fluid 26 remains in oropharynx 30 of the subject. In general, the subject tilts his or her head back during all of the gargling and/or swishing steps of the methods described and claimed herein. As used in the present application, including in the claims and Inventive Concepts, “gargling” means holding a liquid in the throat (typically in the oropharynx) and agitating with air expelled from the lungs. In general, during gargling, the subject avoids swallowing to the extent reasonably possible.

[0475] Reference is still made to FIGS. 2A-D and is additionally made to FIGS. 5A-B, which are schematic illustrations of a portion of a method for collecting specimen sample 24 from the subject, in accordance with an application of the present invention. In some applications, the method of FIGS. 2A-D further comprises, before collecting specimen sample 24, orally dispensing oral wash fluid 68 into oral cavity 36 via anterior opening 34 (typically by the subject), such as shown in FIG. 5A, before or after intranasally introducing nasal wash fluid 26, to combine with the intranasally-introduced nasal wash fluid 26 to form a mixture 70 in oropharynx 30, such as shown in FIG. 5B. Thereafter, the subject gargles mixture 70, and specimen sample 24 is collected from oral cavity 36. Specimen sample 24 contains at least a portion of mixture 70 and the at least a portion of the biological material washed into oropharynx 30 with nasal wash fluid 26. Optionally, the subject additionally swishes mixture 70 in oral cavity 36.

[0476] In some applications, the method of FIGS. 2A-D further comprises, before collecting specimen sample 24, orally dispensing oral wash fluid 68 into oral cavity 36 via anterior opening 34 (typically by the subject), such as shown in FIG. 5A, before or after intranasally introducing nasal wash fluid 26. The subject gargles oral wash fluid 68 such that oral wash fluid 68 forms mixture 70 with the intranasally-introduced nasal wash fluid 26 in oropharynx 30 and/or oral cavity 36, such as shown in FIG. 5B. Specimen sample 24 contains at least a portion of mixture 70 and the at least a portion of the biological material washed into oropharynx 30 with nasal wash fluid 26. Optionally, the subject additionally swishes mixture 70 in oral cavity 36.

[0477] In some applications, the method of FIGS. 2A-D further comprises, before collecting specimen sample 24, orally dispensing oral wash fluid 68 into oral cavity 36 via anterior opening 34 (typically by the subject), such as shown in FIG. 5A, before or after intranasally introducing nasal wash fluid 26. The subject gargles oral wash fluid 68. Specimen sample 24 contains at least a portion of oral wash fluid 68, the at least a portion of the biological material washed into the oropharynx with the nasal wash fluid, and typically, but not necessarily, a portion of nasal wash fluid 26. Optionally, the subject additionally swishes oral wash fluid 68 in oral cavity 36.

[0478] In some applications, the method of FIGS. 2A-D further comprises, before collecting specimen sample 24, orally dispensing oral wash fluid 68 into oral cavity 36 via anterior opening 34 (typically by the subject), such as shown in FIG. 5A, before or after intranasally introducing nasal wash fluid 26. The subject swishes oral wash fluid 68 in oral cavity 36. Specimen sample 24 contains at least a portion of oral wash fluid 68, the at least a portion of the biological material washed into the oropharynx with the nasal wash fluid, and typically, but not necessarily, a portion of nasal wash fluid 26.

[0479] In some applications, collecting specimen sample 24 comprises collecting specimen sample 24 from oral cavity 36 of the subject by the subject expressing, e.g., spitting, out the contents of oral cavity 36 from oral cavity 36 via anterior opening 34. For example, the subject may spit out at least a portion of the intranasally-introduced nasal wash fluid 26 and at least a portion of orally-introduced oral wash fluid 68 from oral cavity 36.

[0480] Experiment 8, described hereinbelow, provides evidence of the efficacy of these techniques for collecting specimen samples for detection of SARS-CoV-2 using quantitative PCR (qPCR).

[0481] For example, 5-10 mL of oral wash fluid 68 may be orally dispensed into oral cavity 36 via anterior opening 34.

[0482] In some applications, the subject alternates multiple times between gargling and sniffing or snorting, before collecting specimen sample 24 from oral cavity 36. Typically, the alternation between gargling and sniffing or snorting occurs while nasal wash fluid 26 and/or oral wash fluid 68 remains in oropharynx 30 of the subject.

[0483] For any of the applications described herein, orally dispensing oral wash fluid 68 into oral cavity 36 via anterior opening 34 of oral cavity 36 may comprise orally dispensing oral wash fluid 68 into oral cavity 36 via anterior opening 34 of oral cavity 36 while the subject is sitting or standing in a vertical position.

[0484] In some applications, oral wash fluid 68 comprises a non-irritant solution; for example, the non-irritant solution may comprise or consist of water. In some applications, the non-irritant solution comprises saline solution that may be hypertonic, isotonic, or hypotonic, for example, a phosphate-buffered saline solution.

[0485] Typically, oral wash fluid 68 does not comprise a drug molecule. Alternatively, oral wash fluid 68 comprises a drug molecule.

[0486] In some applications, oral wash fluid 68 comprises a tastant, for example, menthol flavoring. In some applications, nasal wash fluid 26 and oral wash fluid 68 contain different chemical components that, when combined, generate an indicator. For example, the indicator may be bubbling that is produced by a chemical reaction generated by combining the different chemical components of nasal wash fluid 26 and oral wash fluid 68.

[0487] In some applications, nasal wash fluid 26 comprises a first color additive, and oral wash fluid 68 comprises a second colored additive, and when combined, a third color is generated, for example, by mixing of the two color additives, or for example, by a chemical reaction. In some applications, collecting specimen sample 24 comprises confirming that specimen sample 24 contains at least a portion of nasal wash fluid 26 and at least a portion of oral wash fluid 68 by visually ascertaining that specimen sample 24 has an indicator produced by a combination of nasal wash fluid 26 and oral wash fluid 68, for example, a third color produced by a combination of the first and the second color additives. For some applications, visually ascertaining is performed by the human eye. Alternatively, for some applications, visually ascertaining is performed using spectroscopy, image processing, and/or three-dimensional (3D) image sensing; any of these techniques may optionally utilize artificial intelligence.

[0488] Alternatively or additionally, in some applications, oral wash fluid 68 comprises a chemical that will increase the amount of biological material in specimen sample 24 by enhancing the clearance of the mucus as well as dislodging the biological material lining the mucosa, for example, a mucolytic agent such as acetylcysteine, or for example, a detergent such as polysorbate, or for example, a gas such as pressurized carbon dioxide to generate bubbles.

[0489] Alternatively, in some applications, collecting specimen sample 24 comprises collecting a specimen sample from oral cavity 36 of the subject by aspirating out (using suction) at least a portion of nasal wash fluid 26 and at least a portion of the biological material from oral cavity 36 via anterior opening 34, for example, using a catheter. For example, the at least a portion of the intranasally-introduced nasal wash fluid and orally introduced oral wash fluid 68 may be aspirated out from oral cavity 36 after gargling by the subject. Alternatively or additionally, in some applications, collecting a specimen sample comprises collecting a specimen sample from oral cavity 36 of the subject by drawing out at least a portion of nasal wash fluid 26 and at least a portion of the biological material from oral cavity 36, using absorbent material. For example, the at least a portion of the intranasally-introduced nasal wash fluid and orally introduced oral wash fluid 68 may be drawn from oral cavity 36 using absorbent material after gargling by the subject.

[0490] Gargling nasal wash fluid 26 or mixture 70 of nasal wash fluid 26 and oral wash fluid 68 predominately bathes the anterior oropharynx, including the tonsils, tonsillar pillars, uvula 69, soft palate 71, and posterior tongue 73. In this application, the method thus allows at least a portion of the intranasally-introduced nasal wash fluid that sampled nasal cavity 22 and nasopharynx 32 to also sample oropharynx 30 via gargling and to then be collected from oral cavity 36. Additionally, gargling may also allow for collection of respiratory particles and droplets expressed from or found on posterior pharynx 42, hypopharynx 44, oral cavity 36, and lower airways since the act of gargling is a process of exhaling. Therefore, specimen sample 24 collected by this method may contain droplets and particles from the entire respiratory tract including both upper and lower airways. Typically, the collected specimen sample contains at least portion of the nasally-introduced nasal wash fluid solution and at least a portion of the orally introduced oral wash fluid 68 solution and at least a portion of the biological material that originated from the upper aerodigestive tract. Alternatively, the collected specimen sample contains at least portion of the nasally-introduced nasal wash fluid solution and at least a portion of the biological material that originated from the upper aerodigestive tract.

[0491] In some applications, the biological material that originated from the upper aerodigestive tract includes any of the biological material listed hereinabove with reference to FIGS. 2A-D. multiple biological materials or particulates that may include: microorganisms, antigens, human cells, extracellular soluble and non-soluble particles, cellular components and products, blood products cellular biomarkers, hormones, vitamins, electrolytes, chemical mediators from cells such as mediators of inflammation, pollens, mucus, saliva, sputum, respiratory particles, droplets derived from the upper and lower airways, nucleic acids including DNA and RNA, and chemicals originating from external vapors. Typically, microorganisms collected in specimen sample 24 may include either pathogenic microorganisms or non-pathogenic microorganisms or both, for example, viruses, bacteria, protozoa, and fungi. Typically, human cells collected in specimen sample 24 may include epithelial cells, for example, columnar epithelial cells primarily derived from the nasal cavity and squamous epithelial cells primarily derived from the oral cavity. Typically, human cells collected in specimen sample 24 may also include cellular responders of the immune system, for example, neutrophils, eosinophils, lymphocytes, monocytes, macrophages, mast cells, and histocytes.

[0492] For some applications, the method further comprises waiting for a period of time between (a) concluding intranasally dispensing nasal wash fluid 26 into nasal cavity 22 and (b) orally dispensing oral wash fluid 68 into oral cavity 36, the period of time at least 1 minute, e.g., at least 2 minutes, at least 3 minutes, or at least 5 minutes, such as at least 10 minutes. Optionally, the period of time is at least 15 minutes, or at least 30 minutes. The inventors found that waiting for a period of time less than 90 minutes between intranasally dispensing nasal wash fluid 26 into nasal cavity 22 and orally dispensing oral wash fluid 68 into oral cavity 36 increased the quantity of virus in the collected sample.

[0493] For some applications, specimen sample 24 is a first specimen sample, and mixture 70 is a first mixture, and the method further comprises, after collecting the first specimen sample, waiting for a period of time, the period of time at least 1 minute, e.g., at least 2 minutes, at least 3 minutes, or at least 5 minutes, such as at least 10 minutes. Optionally, the period of time is at least 15 minutes, or at least 30 minutes. After waiting, additional oral wash fluid 68 is orally dispensed into oral cavity 36 via anterior opening 34 of oral cavity 36 such that the additional oral wash fluid 68 forms a second mixture with nasal wash fluid 26 in oropharynx 30. Thereafter, the subject gargles the second mixture. Thereafter, a second specimen sample is collected that passed out of anterior opening 34 of oral cavity 36 and contains at least a portion of the second mixture and the at least a portion of the biological material washed into oropharynx 30 with nasal wash fluid 26. The method described in this paragraph typically does not comprise intranasally dispensing nasal wash fluid into nasal cavity 22 after collecting the first specimen sample. Optionally, the period of time is at least 15 minutes, or at least 30 minutes. The inventors found that virus was detected in both the first and second specimen samples, even though additional nasal wash was not intranasally dispensed before the additional oral wash fluid 68 was dispensed and gargled.

[0494] Reference is again made to FIG. 2D. In some applications, collecting the one or more specimen samples 24 comprises collecting one or more fluid specimen samples 24. In some applications, collecting one or more specimen samples 24 consists collecting at least two specimen samples, for example, collecting exactly two specimen samples. In some applications, collecting the at least two specimen samples comprises collecting further specimen samples in addition to collecting specimen sample 24 from oral cavity 36, for example, collecting an additional specimen sample by swabbing nasal cavity 22 or by collecting an additional specimen sample by inserting absorbent material into nasal cavity 22, typically after collecting fluid specimen sample 24 from oral cavity 36. In some applications, first specimen sample 24 contains at least a first portion of nasal wash fluid 26 and at least a first portion of the biological material, for example collected from oral cavity 36, and a second specimen sample contains at least a second portion of the portion of nasal wash fluid 26 and at least a second portion of the biological material, for example collected from nasal cavity 22.

[0495] In some applications, the two or more specimen samples are combined to produce a combined specimen sample.

[0496] In some applications, collecting the second specimen sample comprises collecting from nasal cavity 22 by the subject blowing specimen sample 24 out of nasal cavity 22 via the one or two nostrils 28. In some applications, collecting the second specimen sample comprises collecting the second specimen sample that was blown, by the subject, out of nasal cavity 22 via the one or two nostrils 28.

[0497] In other applications, collecting the second specimen sample comprises collecting from nasal cavity 22 by aspirating specimen sample 24 out of nasal cavity 22 via the one or two nostrils 28, for example, using a catheter.

[0498] In some applications, collecting the second specimen sample by aspirating specimen sample 24 out from nasal cavity 22 via the one or two nostrils 28 comprises using a negative pressure vacuum generated by nasal wash fluid dispenser 20 after releasing nasal wash fluid 26, for example, by using nasal wash fluid dispenser 20 in accordance with an application of the present invention.

[0499] In some applications, collecting a second specimen sample from nasal cavity 22 comprises, after collecting a specimen sample from oral cavity 36 in collection receptacle 40, collecting residual biological material and nasal wash fluid 26 from nasal cavity 22 by blowing out of the nose of the subject into a plastic bag that is attached to collection receptacle 40. For example, the plastic bag may have a distal opening that is hermetically sealed to the proximal opening of collection receptacle 40 and may have a proximal opening that has a sealable closure mechanism, to allow for specimen sample 24 blown out of the nose of the subject into the plastic bag to join with specimen sample 24 collected from oral cavity 36 in collection receptacle 40. There may be a sealable opening or spout in collection receptacle 40 or in the plastic bag that is openable to allow exit of the combined specimen sample for testing.

[0500] Reference is again made to FIG. 2B. In some applications, instead of or in addition to intranasally dispensing nasal wash fluid 26 into nasal cavity 22, a vapor (e.g., a warm vapor) is intranasally dispensed into nasal cavity 22 to loosen the biological material in nasal cavity 22. Typically, intranasally dispensing the vapor comprises inhaling, by the subject, the vapor into nasal cavity 22, and, typically, into oropharynx 30 via nasopharynx 32, such that the vapor washes the biological material into oropharynx 30 from (a) nasal cavity 22, (b) nasopharynx 32, or (c) nasal cavity 22 and nasopharynx 32. Typically, specimen sample 24 contains at least a portion of the biological material washed into oropharynx 30 by the vapor.

[0501] Reference is made to FIGS. 2A-5B. The sampling methods described herein with reference to FIGS. 2A-5B typically do not comprise swabbing any portion of the subject, unless the methods are specifically described as comprising swabbing a portion of the subject. Alternatively, the sampling methods described herein may optionally comprise swabbing a portion of the subject.

[0502] The sampling methods described herein with reference to FIGS. 2A-5B typically do not comprise swabbing nasal cavity 22, unless the methods are specifically described as comprising swabbing nasal cavity 22. Alternatively, the sampling methods described herein may optionally comprise swabbing nasal cavity 22.

[0503] The sampling methods described herein with reference to FIGS. 2A-5B typically do not comprise swabbing nasopharynx 32, unless the methods are specifically described as comprising swabbing nasopharynx 32. Alternatively, the sampling methods described herein may optionally further comprise swabbing nasopharynx 32.

[0504] In some applications of the present invention, the methods with reference to FIGS. 2A-5B do not comprise swabbing nasopharynx 32, and specimen sample 24 is collected by swabbing oropharynx 30. For some of these applications, the methods do not comprise swabbing any portion of nasal cavity 22. Alternatively, for some of these applications, collecting specimen sample 24 comprises swabbing oropharynx 30 and swabbing one or both anterior nares of nasal cavity 22. Alternatively or additionally, for some of these applications, collecting specimen sample 24 comprises swabbing oropharynx 30 and swabbing one or more turbinates of nasal cavity 22 selected from the group consisting of: a middle turbinate and an inferior turbinate. As is known in the art, the anterior naris is the portion of the nasal cavity anterior to the internal nasal valve.

[0505] The sampling methods described herein with reference to FIGS. 2A-5B typically do not comprise swabbing oropharynx 30 and do not comprise swabbing nasopharynx 32, unless the methods are specifically described as comprising swabbing oropharynx 30 and/or nasopharynx 32. Alternatively, the sampling methods described herein may optionally further comprise swabbing oropharynx 30 and/or nasopharynx 32.

[0506] For some applications, collecting specimen sample 24 comprises drawing specimen sample 24 out of oral cavity 36 via anterior opening 34 of oral cavity 36 using an absorbent material (e.g., a flocked or cotton swab, or a sponge). (For example, an ORAcollect®⋅RNA Saliva Collection Device (DNA Genotek Inc., a subsidiary of OraSure Technologies, Inc. (Bethlehem, Pa., USA)) may be used.) Optionally, the absorbent material is located on a tip of a collector shaft, and specimen sample 24 is drawn out of oral cavity 36 via anterior opening 34 of oral cavity 36 using the absorbent material by inserting the tip of the collector shaft into oral cavity 36. For some of these applications, specimen sample 24 is drawn out of oral cavity 36 via anterior opening 34 of oral cavity 36 using the absorbent material by the subject sucking on the absorbent material.

[0507] For some applications in which the method does not comprise swabbing oropharynx 30 and does not comprise swabbing nasopharynx 32, specimen sample 24 is collected by drawing specimen sample 24 out of oral cavity 36 via anterior opening 34 of oral cavity 36 by contacting one or more portions of oral cavity 36 with an absorbent material (e.g., at a tip of a collector shaft), without swabbing oropharynx 30. (For example, the above-mentioned ORAcollect®⋅RNA Saliva Collection Device may be used.) For example, the one or more portions of oral cavity 36 may include one or more of buccal mucosa, the tongue (e.g., under the tongue), the gums (e.g., the lower gums), and/or the palatal mucosa. For example, for swabbing the lower gums, absorbent material (e.g., at a tip of a collector shaft) may be rubbed back and forth along the lower gums several times.

[0508] In applications of the present invention that comprise swabbing the subject, specimen sample 24 may be collected into collection receptacle 40 by placing the swab (or swabs) in collection receptacle 40.

[0509] In some applications, the methods described comprise testing the one or more specimen samples for the presence of a particulate, for example, testing the combined specimen sample for the presence of a particulate. Typically, but not necessarily, the particulate is present in at least the biological material washed from nasal cavity 22 and/or nasopharynx 32 to oropharynx 30, and, optionally, also in other biological material from elsewhere in the respiratory tract.

[0510] In some applications, testing the one or more specimen samples for the presence of a particulate comprises first concentrating specimen sample 24 by filtration prior to testing for the presence of a particulate (by passing specimen sample 24 through one or more filters).

[0511] In some applications, testing the one or more specimen samples for the presence of a particulate comprises testing to aid in the diagnosis of disease, for example, aiding in the diagnosis of infectious diseases or allergies. In some applications, testing the one or more specimen samples for the presence of a particulate comprises testing to aid in the diagnosis of a biological status or inflammatory status. In some applications, testing for the presence of a particulate comprises testing for the presence of one or more of the following soluble or insoluble particulates: a pathogenic microorganism, a non-pathogenic microorganism, an antigen, a human cell, a cellular biomarker, a hormone, a chemical mediator from cells such as a mediator of inflammation, a pollen, a respiratory particle, a particle contained within droplets from the lower airways of the subject, a nucleic acid including DNA and RNA, and a chemical originating from an external vapor. In some applications, the testing for the presence of a particulate comprises testing for the presence of one or more of the following microorganisms: viruses, bacteria, and fungi, e.g., testing for the presence of bacteria, for example, Streptococcus bacteria, such as Streptococcus pyogenes (Strep A), or, for example, testing for the presence of viruses such as Influenza viruses, or for example, coronaviruses, such as SARS-CoV-2. In some applications, specimen sample 24 is tested for the presence of cerebrospinal fluid (CSF).

[0512] In some applications, the testing for the presence of a particulate comprises testing for the presence of one or more of the following antigens: viral antigens, bacterial antigens, and fungal antigens.

[0513] In some applications, the testing for the presence of a particulate comprises microscopic imaging, fluorescence activated cell sorting (FACS), spectroscopy, microscopy (e.g., automated digital microscopy), image processing, three-dimensional (3D) image sensing, and/or CRISPR diagnostic testing.

[0514] In some applications, the testing for the presence of a particulate comprises artificial intelligence. Any of the above-mentioned testing techniques may optionally utilize artificial intelligence.

[0515] In some applications, the testing for the presence of a particulate comprises culturing at least a portion of specimen sample 24.

[0516] In some applications, the testing for the presence of a particulate comprises performing a particulate detection test. In some applications, the particulate detection test is an immunoassay, for example, a lateral-flow immunochromatographic assay, a chromatographic digital immunoassay, or an enzyme-linked immunosorbent assay (ELISA). In some applications, the particulate detection test is a molecular-based assay, for example, nucleic acid hybridization, or for example, nucleic acid amplification, including polymerase chain reaction (PCR) amplification, real-time quantitative PCR (qPCR) amplification, reverse transcriptase PCR (RT-PCR) amplification, and isothermal amplification, such as loop-mediated isothermal amplification (LAMP). For some of these applications, specimen sample 24 is transported from the site of collection to a remote site, such as a remote laboratory, for testing. For these applications, specimen sample 24, or a portion thereof, may be stored in a liquid, such as a lysis buffer, saline solution (e.g., phosphate buffered saline (PBS)), transport medium (e.g., universal transport medium or a viral transport medium), or a composition comprising a detergent and a buffering agent for stabilizing RNA (e.g., distributed by DNA Genotek Inc., a subsidiary of OraSure Technologies, Inc. (Bethlehem, Pa., USA), for example, ORAcollect®⋅RNA Saliva Collection Device for SARS CoV-2). The liquid is optionally contained in collection receptacle 40, described hereinabove with reference to FIG. 2D.

[0517] In some applications, the testing for the presence of the particulate comprises performing a pretesting step prior to testing for the presence of the particulate, comprising one or more of the following pretesting steps: chemical treatment, purification, filtration, enrichment, and/or centrifugation. In other applications, specimen sample 24 is tested directly for the presence of a particulate without any pretesting steps.

[0518] Reference is again made to FIGS. 2A-D. In some applications of the present invention, a method is provided that comprises intranasally dispensing nasal wash fluid into nasal cavity 22 such that nasal wash fluid 26 passes from nasal cavity 22 into oropharynx 30 via nasopharynx 32. The method further comprises, thereafter, collecting a specimen sample that passed out of anterior opening 34 of oral cavity 36 and contains at least a portion of nasal wash fluid 26. In this method, biological material of interest for subsequent testing is not necessarily washed from nasal cavity 22 or nasopharynx 32 into oropharynx 30, although, of course, some biological material, such as mucus, will be thus washed. Instead, the biological material of interest for subsequent testing comes from elsewhere in the respiratory tract, typically at least oropharynx 30, and optionally lower in the respiratory tract. In this method, nasal wash fluid 26, upon arriving in oropharynx 30, may help loosen and increase the collection of biological material from the walls (e.g., of posterior pharynx 42) of oropharynx 30, and/or may provide better coverage of posterior pharynx 42 than gargling alone, and thus result in better sampling of posterior pharynx 42 than gargling alone. Thus, this method may be useful for detecting particulates present in the respiratory tract only at the level of oropharynx 30 or lower, but not in nasal cavity 22 or nasopharynx 32.

[0519] This method may be practiced in combination with the other methods described herein, including, but not limited to gargling and/or spraying, via anterior opening 34 of oral cavity 36, oropharynx 30 with oropharyngeal fluid wash.

[0520] In some applications, specimen samples 24 collected using the collection techniques described herein are tested for a particulate using one or more of the techniques described in the patent applications incorporated hereinbelow by reference, or a technique known in the art. The particulate may be a virus, a bacterium, any of the other particulates described hereinabove, or any of the other biological materials described hereinabove.

[0521] Reference is again made to FIG. 4. In some applications, a method is provided that comprises spraying (either by a healthcare worker or the subject), via anterior opening 34 of oral cavity 36, oropharynx 30 with oropharyngeal fluid wash 46. Thereafter, a specimen sample is collected that passed out of anterior opening 34 of oral cavity 36 (e.g., was spit out of anterior opening 34) and contains at least a portion of biological material washed off a pharyngeal wall (typically, the posterior pharyngeal wall) into oropharynx 30 of the subject by the oropharyngeal fluid wash. Specimen sample 24 may additionally or alternatively contain at least a portion of biological material washed off one or more of the following locations: posterior pharynx 42, a tonsillar area, soft palate 71, and posterior tongue 73. This method may be performed without intranasally dispensing nasal wash fluid into nasal cavity 22, or it may be performed in combination with such dispensing, such as described hereinabove. This method may be performed in combination with any of the techniques described herein, such as the particulate testing techniques described herein. For some applications, a spray bottle 47 is provided that contains oropharyngeal fluid wash 46; spray bottle 47 may be an element of a kit comprising other elements described herein.

[0522] In some applications, the method further comprises, before collecting specimen sample 24, gargling the oropharyngeal fluid wash by the subject. This method may produce a better specimen sample than would be produced by gargling alone, because gargling alone generally results in the gargle fluid having limited contact with the posterior pharyngeal wall. Alternatively, the method does not comprise gargling the oropharyngeal fluid wash by the subject. Further alternatively or additionally, in some applications, the method further comprises, before collecting specimen sample 24, swishing, by the subject, the oropharyngeal fluid wash in oral cavity 36.

[0523] In some applications, the method further comprises, before collecting specimen sample 24, orally dispensing oral wash fluid 68 into oral cavity 36 via anterior opening 34 of oral cavity 36 such that oral wash fluid 68 forms a mixture with the oropharyngeal fluid wash in oropharynx 30; and gargling and/or swishing mixture 70 by the subject. In these applications, specimen sample 24 contains at least a portion of mixture 70 and the at least a portion of the biological material washed off the pharyngeal wall into oropharynx 30 by mixture 70.

[0524] In some applications, the spraying is performed while the subject says “ah” or “eh.” This may increase the amount of the oropharyngeal fluid wash that is deposited on the posterior pharyngeal wall, such as to build up enough volume to gargle and/or swish.

[0525] In some applications, the biological material includes one or more particulates selected from the group consisting of: a virus (e.g., a coronavirus, such as SARS-CoV-2; or an Influenza virus), a bacterium (e.g., Streptococcus bacterium), a protozoan, and a fungus.

[0526] In some applications, a total volume of at least 2 mL (e.g. 2-15 mL, e.g. 2-10 mL) of the oropharyngeal fluid wash is sprayed.

[0527] In some applications, oropharynx 30 is sprayed with the oropharyngeal fluid wash in a plurality of pulses. For example, each of the pulses may have a volume of 0.05-5 mL.

[0528] In some applications, collecting specimen sample 24 comprises expressing (e.g., spitting), by the subject, specimen sample 24 out of oropharynx 30 via the anterior opening of oral cavity 36.

[0529] In some applications of the present application, a method is provided for testing for the presence of a particulate. The particulate may be a virus, a bacterium, any of the other particulates described hereinabove, or any of the other biological materials described hereinabove. This method may optionally be combined with any of the techniques described herein, including the techniques for intranasally dispensing nasal wash fluid. Alternatively, this method may be practiced separately from the other techniques described herein, including without intranasally dispensing nasal wash fluid.

[0530] The method comprises collecting, from a subject, a liquid specimen sample that that includes orally-dispensed oral wash fluid 68 and potentially contains the particulate. For some applications, the particulate is selected from the group consisting of: a virus, a bacterium, a protozoan, and a fungus. For example, the particulate may be an Influenza virus or a coronavirus, such as Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Alternatively or additionally, the particulate may be an antibody, an antigen, or a leukocyte, or any of the other particulates described herein, and/or the liquid specimen sample potentially contains any of the biological materials described herein.

[0531] The liquid specimen sample is passed through one or more porous filters to produce a filtrate. For some applications, the one or more porous filters comprise at least one filter having an average absolute or nominal pore size of between 0.1 and 3 microns, such as between 0.2 and 3 microns, such as between 0.2 and 2 microns, e.g., between 0.45 and 2 microns, such as between 0.8 and 2 microns, e.g., between 1 and 2 microns, such as between 1.2 and 2 microns. Alternatively, the average absolute or nominal pore size may be 3-6 microns, 2-10 microns, 6-13 microns, 10-20 microns, 13-20 microns, or 20-40 microns. Alternatively, the average absolute or nominal pore size may be between 0.03 and 0.3 microns. Further alternatively or additionally, the at least one filter may have the pore sizes described hereinbelow with reference to FIGS. 14A-B for large-pored filter(s) 520 of filter units 500A and 500B. Optionally, the above-mentioned at least one filter is at least one first filter, and the one or more porous filters comprise a second filter, downstream from the at least one first filter, the second filter having an average absolute or nominal pore size less than that of the at least one first filter. For example, the average absolute or nominal pore size of the second filter may be between 0.03 and 0.3 microns.

[0532] The filtrate is tested for the presence of the particulate (or the other biological materials described herein).

[0533] For some applications, collecting the liquid specimen sample comprises: [0534] gargling, by the subject, oral wash fluid 68 to produce the liquid specimen sample that includes oral wash fluid 68 and potentially contains the particulate; and [0535] thereafter, collecting the liquid specimen sample from the subject.

[0536] For some applications, collecting the liquid specimen sample comprises: [0537] swishing, by the subject, oral wash fluid 68 in oral cavity 36 of the subject to produce the liquid specimen sample that includes the oral wash fluid and potentially contains the particulate; and [0538] thereafter, collecting the liquid specimen sample from the subject.

[0539] For some applications, collecting the liquid specimen sample comprises: [0540] spraying, via anterior opening 34 of oral cavity 36, oropharynx 30 with oral wash fluid 68; and [0541] thereafter, collecting the liquid specimen sample from the subject.

[0542] For some applications, testing for the presence of the particulate comprises using spectroscopy to test for the presence of the particulate, using microscopy (e.g., automated digital microscopy) to test for the presence of the particulate, using image processing to test for the presence of the particulate, and/or using three-dimensional (3D) image sensing to test for the presence of the particulate. For some applications, testing for the presence of the particulate comprises using a molecular-based assay (such as nucleic acid amplification, e.g. polymerase chain reaction (PCR) or isothermal amplification, e.g., loop-mediated isothermal amplification (LAMP)) to test for the presence of the particulate. For some applications, testing for the presence of the particulate comprises using another testing technique to test for the presence of the particulate, such as any of the testing techniques described herein. For some applications, testing for the presence of the particulate comprises using artificial intelligence to test for the presence of the particulate. Any of the above-mentioned testing techniques may optionally utilize artificial intelligence.

[0543] Typically, testing the filtrate for the presence of the particulate comprises diagnosing a disease.

[0544] Typically, the method does not comprise centrifuging the liquid specimen sample before passing the liquid specimen sample through the one or more porous filters.

[0545] Typically, the method does not comprise centrifuging the filtrate before testing the filtrate for the presence of the particulate.

[0546] Alternatively, the method comprises centrifuging the liquid specimen sample before passing the liquid specimen sample through the one or more porous filters, and/or centrifuging the filtrate before testing the filtrate for the presence of the particulate.

[0547] Typically, the method does not comprise culturing the liquid specimen sample before passing the liquid specimen sample through the one or more porous filters.

[0548] Typically, the method does not comprise culturing the filtrate before testing the filtrate for the presence of the particulate.

[0549] Alternatively, the method comprises culturing the liquid specimen sample before passing the liquid specimen sample through the one or more porous filters, and/or culturing the filtrate before testing the filtrate for the presence of the particulate.

[0550] In an application of the present invention, a method is provided for testing for the presence of a particulate. The particulate may be a virus, a bacterium, any of the other particulates described hereinabove, or any of the other biological materials described hereinabove. This method may optionally be combined with any of the techniques described herein, including the techniques for intranasally dispensing nasal wash fluid. Alternatively, this method may be practiced separately from the other techniques described herein, including without intranasally dispensing nasal wash fluid.

[0551] The method comprises collecting, from a subject, a liquid specimen sample that that includes orally-dispensed oral wash fluid 68 and potentially contains a particulate selected from the group consisting of: a virus, a bacterium, a protozoan, and a fungus. For example, the particulate may be an Influenza virus or a coronavirus. such as Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Alternatively or additionally, the particulate may be an antibody, an antigen, or a leukocyte, or any of the other particulates described herein, and/or the liquid specimen sample potentially contains any of the biological materials described herein.

[0552] The liquid specimen sample is tested for the presence of the particulate using spectroscopy, microscopy (e.g., automated digital microscopy), image processing, three-dimensional (3D) image sensing, and/or artificial intelligence.

[0553] For some applications, collecting the liquid specimen sample comprises: [0554] gargling, by the subject, oral wash fluid 68 to produce the liquid specimen sample that includes oral wash fluid 68 and potentially contains the particulate; and [0555] thereafter, collecting the liquid specimen sample from the subject.

[0556] For some applications, collecting the liquid specimen sample comprises: [0557] swishing, by the subject, oral wash fluid 68 in oral cavity 36 of the subject to produce the liquid specimen sample that includes the oral wash fluid and potentially contains the particulate; and [0558] thereafter, collecting the liquid specimen sample from the subject.

[0559] For some applications, collecting the liquid specimen sample comprises: [0560] spraying, via anterior opening 34 of oral cavity 36, oropharynx 30 with oral wash fluid 68; and [0561] thereafter, collecting the liquid specimen sample from the subject.

[0562] For some applications, the liquid specimen sample is passed through one or more porous filters to produce a filtrate, and the filtrate is tested for the presence of the particulate. For example, the one or more filters may have any of the pore sized described hereinabove for the previous method described immediately above.

[0563] For some applications, the liquid specimen sample is concentrated by filtration prior to testing for the presence of the particulate, by passing the liquid specimen sample through one or more filters. For example, the one or more filters may have any of the pore sized described hereinabove for the previous method described immediately above. For example, at least one of the one or more filters may have a pore size of 0.03 to 0.3 microns, and/or the pore sizes described hereinbelow with reference to FIGS. 14A-B for large-pored filter(s) 520 of filter units 500A and 500B. Optionally, the liquid specimen sample is tested for the presence of the particulate by analyzing the one or more filters using spectroscopy, microscopy (e.g., automated digital microscopy), image processing, three-dimensional (3D) image sensing, and/or artificial intelligence.

[0564] Typically, testing for the presence of the particulate comprises diagnosing a disease.

[0565] Typically, the method does not comprise centrifuging the liquid specimen sample before testing for the presence of the particulate.

[0566] Typically, when the particulate is a virus, the method does not comprise culturing the liquid specimen sample before testing for the presence of the particulate. Alternatively, for some applications, such as when the particulate is a bacterium, the method further comprises culturing the liquid specimen sample before testing for the presence of the particulate.

[0567] Alternatively, the method comprises centrifuging the liquid specimen sample before testing for the presence of the particulate, and/or culturing the liquid specimen sample before testing for the presence of the particulate.

[0568] Reference is again made to FIGS. 1, 2A-D, and 3A-B, and is additionally made to FIG. 6, which is a schematic illustration of nasal wash fluid dispenser 20 with tubular applicator 64 inserted into nostril 28, in accordance with an application of the present invention. Reference is also made to FIGS. 7A-K, which are several schematic illustrations of nasal wash fluid dispenser 20 viewed from several different directions, in accordance with an application of the present invention.

[0569] In some applications of the present invention, nasal wash fluid dispenser 20 comprises a dispensing container 72, which contains nasal wash fluid 26; and tubular applicator 64, which extends distally from and is in fluid communication with dispensing container 72. Tubular applicator 64 is sized and shaped to be insertable into nostril 28.

[0570] The tubular applicator 64 comprises first and second nozzles 60A, 60B, which are configured to dispense nasal wash fluid 26 in first and second fluid discharges, respectively, having first and second centerlines 74A and 74B (labeled in FIG. 3A), respectively, first and second centerlines 74A and 74B defining an offset angle α (alpha) (labeled in FIG. 3A) therebetween of between 10 and 90 degrees (such as between 20 and 80 degrees, e.g., between 60 and 70 degrees, such as between 30 and 60 degrees, e.g., between 40 and 50 degrees). As a result, when tubular applicator 64 is inserted into nostril 28 with first nozzle 60A oriented in posterior direction 48 and second nozzle 60B oriented in superoposterior direction 50, as shown in FIG. 3A, first nozzle 60A directs the first fluid discharge toward nasopharynx 32 and second nozzle 60B directs the second fluid discharge toward olfactory area 52 of nasal cavity 22, such that nasal wash fluid 26 washes biological material into oropharynx 30 from (a) nasal cavity 22, (b) nasopharynx 32, or (c) nasal cavity 22 and nasopharynx 32. First nozzle 60A directs at least a portion of the first fluid discharge under an inferior turbinate 76, so as to reach toward nasopharynx 32.

[0571] In some applications, first and second nozzles 60A, 60B are configured to dispense the first and the second fluid discharges of nasal wash fluid 26 with the same volume, sprayed particle size, velocity, and flow rate.

[0572] In some applications, first and second nozzles 60A, 60B are configured to dispense the first and the second fluid discharges of nasal wash fluid 26 with respective different fluid flow characteristics, the fluid flow characteristics including one or more characteristics selected from the group consisting of: volume, sprayed particle size, velocity, and flow rate.

[0573] In some applications, one of first and second nozzles 60A, 60B is located at a distal end of tubular applicator 64, and the other of first and second nozzles 60A, 60B is located is located away from the distal end of tubular applicator 64. For some of these applications, tubular applicator 64 is shaped so as to define a distal-most central longitudinal axis 78 through a distal end 80 of tubular applicator 64 (labeled in FIG. 6), and the one of first and second nozzles 60A, 60B located at the distal end of tubular applicator 64 is oriented such that the centerline thereof is coaxial with distal-most central longitudinal axis 78.

[0574] In some applications, first nozzle 60A is configured to dispense the first discharge with a first spray angle β (beta) of between 5 and 30 degrees (e.g., between 5 and 15 degrees), measured at 3 mm from first nozzle 60A (labeled in FIG. 6). In some applications, second nozzle 60B is configured to dispense the first discharge with a second spray angle of between 20 and 60 degrees (not labeled).

[0575] In some applications, tubular applicator 64, including first and second nozzles 60A, 60B, is configured to dispense, via first and second nozzles 60A, 60B, respectively, first and second fractions of a total volume of nasal wash fluid 26 dispensed via tubular applicator 64. The second fraction equals at least two times the first fraction, such as at least five times the first fraction, e.g., at least eight times the first fraction.

[0576] In some applications, nasal wash fluid dispenser 20 is configured to dispense nasal wash fluid by manual compression of the fluid dispenser.

[0577] In some applications, nasal wash fluid dispenser 20 comprises an atomizer that is configured to dispense nasal wash fluid 26.

[0578] In some applications, nasal wash fluid dispenser 20 comprises a pump that is configured to dispense nasal wash fluid 26.

[0579] In some applications, nasal wash fluid dispenser 20 comprises a pressurized aerosol generator that is configured to dispense nasal wash fluid 26.

[0580] In some applications, dispensing container 72 and tubular applicator 64 are shaped so as to be attachable to a pressure machine. For example, the pressure machine may be configured to apply pressure to dispensing container 72 and tubular applicator 64 so as to dispense a metered amount of nasal wash fluid 26. Optionally, the pressure machine may be configured to dispense the metered amount of nasal wash fluid 26 at adjustable pressures, doses and/or configurations.

[0581] In some applications, nostril 28 is a first nostril 28, and tubular applicator 64 is a first tubular applicator 64 that is sized and shaped to be insertable into first nostril 28. Nasal wash fluid dispenser 20 further comprises a second tubular applicator (not shown), which extends distally from and is in fluid communication with dispensing container 72, which is sized and shaped to be insertable into a second nostril 28, and which comprises third and fourth nozzles, which are configured to dispense nasal wash fluid 26 in third and fourth fluid discharges, respectively, having third and fourth centerlines, respectively, the third and the fourth centerlines defining the offset angle therebetween of between 10 and 90 degrees, or the other angles mentioned above. The second tubular applicator may implement any of the features of tubular applicator 64 described hereinabove or hereinbelow.

[0582] In some applications, the first tubular applicator does not comprise any nozzles other than first and second nozzles 60A, 60B, the second tubular applicator does not comprise any nozzles other than the third and the fourth nozzles, and nasal wash fluid dispenser 20 does not comprise any nozzles other than the first, the second, the third, and the fourth nozzles. Alternatively, the first tubular applicator, the second tubular applicator, and/or nasal wash fluid dispenser 20 comprises one or more additional nozzles (configuration not shown).

[0583] In some applications, tubular applicator 64 is malleable and compressible to fit into and occlude nostril 28. Alternatively, tubular applicator 64 is rigid.

[0584] In some applications, tubular applicator 64 comprises a curved portion.

[0585] In some applications, tubular applicator 64 is shaped so as to define an applicator centerline that defines a plane, and first and second nozzles 60A, 60B are configured to dispense nasal wash fluid 26 in the first and the second fluid discharges, respectively, such that first and second centerlines 74A, 74B, respectively, fall in the plane.

[0586] In some applications, tubular applicator 64 is shaped so as to define distal-most longitudinal axis 78 and a proximal-most central longitudinal axis 82 (labeled in FIG. 6) through distal end 80 and a proximal end 84 of tubular applicator 64, respectively, and distal-most and proximal-most central longitudinal axes 78, 82 form a bend angle δ (delta) of between 100 and 245 degrees therebetween (labeled in FIG. 6).

[0587] In some applications, nasal wash fluid dispenser 20 is shaped so as to guide introduction of tubular applicator 64 into nostril 28 at a specific orientation with respect to a maxilla 86 of the subject. In some applications, dispensing container 72 of nasal wash fluid dispenser 20 is shaped so as to guide introduction of tubular applicator 64 into nostril 28 at a specific orientation with respect to maxilla 86 of the subject. In other applications, nasal wash fluid dispenser 20 comprises a housing 88 that is shaped so as to guide the introduction of tubular applicator 64 into nostril 28 at the specific orientation with respect to maxilla 86 of the subject (configuration not shown).

[0588] In some applications, such as shown, for example, in FIGS. 3B, 7F, 7G, 7H, and 7I, nasal wash fluid dispenser 20 or dispensing container 72 or housing 88 of nasal wash fluid dispenser 20 is shaped so as to define a concave surface 90 that faces in posterior direction 48 and generally conforms to an outer surface of an upper lip 92 of the subject, and tubular applicator 64 may press the concave surface 90 against upper lip 92 so as to stabilize nasal wash fluid dispenser 20 with respect to maxilla 86. For example, concave surface 90 may have a lateral width W of between 2.5 and 7 cm e.g., 2.5-6 cm, such as 3-5 cm.

[0589] Typically, as shown in FIGS. 2A-B, in a method for using this configuration of nasal wash fluid dispenser 20, inserting tubular applicator 64 into nostril 28 comprises pressing concave surface 90 against upper lip 92 so as to stabilize nasal wash fluid dispenser 20 with respect to maxilla 86.

[0590] In some applications, concave surface 90 is curved. For example, concave surface 90 may be concave about an axis 94 and flat in a direction along axis 94 (labeled in FIG. 7H). Optionally, axis 94 forms an angle θ (theta) of between 75 and 90 degrees with first centerline 74A of the first fluid discharge, such as between 85 and 90 degrees. In some applications, dispensing container 72 is shaped so as to define concave surface 90. In other applications, nasal wash fluid dispenser 20 comprises housing 88 that is shaped so as to define concave surface 90.

[0591] In some applications, first and second nozzles 60A, 60B are configured to dispense nasal wash fluid such that the first and the second fluid discharges, respectively, are respective first and second types of fluid discharges selected from the group consisting of: a fluid stream, a spray, a mist, a pressurized aerosol, steam, and an atomized fluid. The first and the second types of fluid discharges may be the same type or different types.

[0592] In some applications, at least one of first and second nozzles 60A, 60B comprises a fan mechanism configured to dispense nasal wash fluid 26 in a circular rotary motion to generate dispersion within nasal cavity 22.

[0593] In some applications, first and second nozzles 60A, 60B are configured to dispense a total volume of at least 0.2 mL, no more than 20 mL, and/or 0.2-20 mL of nasal wash fluid 26 into nostril 28, such as 0.6-10 mL, e.g., 0.2-10 mL, such as 0.2-5 mL, e.g., 0.6-5 mL of nasal wash fluid 26 into nostril 28.

[0594] In some applications, nasal wash fluid dispenser 20 is configured to dispense nasal wash fluid 26 in a plurality of pulses. For example, nasal wash fluid dispenser 20 may comprise a mechanical pulsator, e.g., comprising one or more rotatable parts, e.g., which are driven to rotate by flow of nasal wash fluid 26 within the dispenser, and/or an electronic pulsator. Alternatively, the user of dispenser 20 may manually dispense nasal wash fluid 26 in pulses by repeatedly activating the dispenser.

[0595] In some applications, the pulses are delivered at intervals of between 0.5 and 2 seconds apart, e.g., 1 second apart.

[0596] In some applications, nasal wash fluid dispenser 20 is configured to dispense nasal wash fluid 26 in a plurality of pulses having a volume of at least 0.02 mL, no more than 10 mL, and/or 0.02-10 mL per pulse, e.g., 0.02-1 mL per pulse.

[0597] In some applications, at least one of first and second nozzles 60A, 60B is configured to dispense nasal wash fluid 26 with a sprayed particle size of 5-50 microns, such as 5-15 microns or 15-20 microns.

[0598] In some applications, such as labeled in FIG. 2A, a nasal wash fluid system 100 is provided that comprises nasal wash fluid dispenser 20, and further comprises nasal wash fluid 26, contained in dispensing container 72.

[0599] In some applications, nasal wash fluid 26 does not comprise a drug molecule. Alternatively, nasal wash fluid 26 does comprise a drug molecule.

[0600] In some applications, nasal wash fluid 26 comprises a non-irritant solution, which may, for example, comprise or consist of water. Alternatively, in some applications, nasal wash fluid 26 of a non-irritant solution consists of a saline solution, such as a phosphate-buffered saline solution.

[0601] In some applications, nasal wash fluid 26 comprises a tastant.

[0602] Alternatively or additionally, in some applications, nasal wash fluid 26 comprises a color additive. For example, the color additive may have a visually-perceptible color that is dominated by a non-white color other than yellow, such as red, green, or blue, which may be easier for a human subject to perceive in a liquid than yellow. Alternatively, the color additive may be yellow.

[0603] Further alternatively or additionally, in some applications, nasal wash fluid 26 comprises a chemical additive, such as described hereinabove with reference to FIGS. 2B and 2C.

[0604] In some applications, nasal wash fluid 26 comprises a first color additive, and nasal wash fluid system 100 further comprises an oral wash fluid 68 comprising a second colored additive, the first and the second color additives having different colors.

[0605] Reference is now made to FIGS. 8A-C, which are schematic illustrations of a nasal wash fluid dispenser 120, in accordance with an application of the present invention. Except as described below, nasal wash fluid dispenser 120 is identical to nasal wash fluid dispenser 20, described hereinabove, and like reference numerals refer to like parts. In nasal wash fluid dispenser 120, at least one of first and second nozzles 60A, 60B comprises a flat nozzle 161 (also known the nozzle art as an “even nozzle”). Optionally, first nozzle 60A comprises a first flat nozzle 161A, and second nozzle 60B comprises a second flat nozzle 161B. For example, the flat nozzles may comprise flat tapered nozzles.

[0606] Reference is now made to FIGS. 9A-F, which are schematic illustrations of a nasal wash fluid dispenser 220, in accordance with an application of the present invention. Reference is further made to FIG. 10, which is a schematic cross-sectional illustration of nasal wash fluid dispenser 220 inserted into nasal cavity 22, in accordance with an application of the present invention. Reference is still further made to FIG. 11, which is a schematic cross-sectional illustration of a tubular applicator 264 of nasal wash fluid dispenser 220, in accordance with an application of the present invention. Except as described below, nasal wash fluid dispenser 220 is identical to nasal wash fluid dispenser 20, described hereinabove, and like reference numerals refer to like parts.

[0607] Tubular applicator 264 comprises, in addition to first and second nozzles 60A, 60B, a third nozzle 260C, which is configured to dispense nasal wash fluid 26 in a third fluid discharge having a third centerline 274C (labeled in FIG. 10). Typically, first and third centerlines 74A and 274C define an offset angle γ (gamma) (labeled in FIG. 10) therebetween of between 75 and 105 degrees (such as between 85 and 95 degrees, e.g., 90 degrees). As a result, when tubular applicator 264 is inserted into nostril 28 with first nozzle 60A oriented in posterior direction 48 and second nozzle 60B oriented in superoposterior direction 50, such as shown in FIG. 10, third nozzle 260C is oriented in superior direction 62, and thus directs the third fluid discharge at least in superior direction 62, toward the inferior border of the upper lateral cartilage of a nasal vestibule 296 of nasal cavity 22 (labeled in FIG. 10). (Nasal vestibule 296 is the portion of nasal cavity 22 anterior to the anterior head of the inferior turbinate.) Typically, third nozzle 260C has a relatively wide third spray angle ε (epsilon) (labeled in FIG. 10), such that third nozzle 260C additionally directs the third fluid discharge bilaterally (i.e., in both lateral directions within the single nasal cavity 22) toward the septum and a lateral wall of nasal cavity 22 (the lateral wall opposite the septum). For example, the third spray angle ε (epsilon) may between 45 and 135 degrees, such as between 60 and 120 degrees, e.g., about 90 degrees, measured at 3 mm from third nozzle 260C. As a result, nasal wash fluid 26 typically additionally washes biological material into oropharynx 30 from the superior wall, septum, and/or lateral wall of nasal cavity 22. More generally, nasal wash fluid 26 typically additionally washes biological material into oropharynx 30 from nasal vestibule 296, which is at least a portion of the area generally sampled by a conventional nasal swab.

[0608] Reference is now made to FIG. 12, which is a schematic illustration of a testing kit 300, in accordance with an application of the present invention. Testing kit 300 comprises nasal wash fluid system 100 (labeled in FIG. 2A), and further comprises a test 302 (e.g., a diagnostic test), which is configured to test for the presence of a particulate in specimen sample 24 collected using the techniques described herein. (As described hereinabove, nasal wash fluid system 100 comprises nasal wash fluid dispenser 20 and nasal wash fluid 26.) Test 302 may optionally be configured to implement testing techniques described herein, in the patent applications incorporated hereinbelow by reference, and/or known in the art. For example, test 302 may further comprise one or more filters configured to concentrate specimen sample 24 by filtration prior to testing for the presence of the particulate. The particulate may be a virus, a bacterium, any of the other particulates described hereinabove, or any of the other biological materials described hereinabove. For some applications, test 302 comprises a lateral flow immunoassay test strip (which may be an element of a chromatographic digital immunoassay, such as the BD Veritor™ system, mentioned immediately below), which is configured to detect the presence of the particulate (such as by detecting protein antigen, e.g., from a virus), and, optionally, one or more reagents. (Lateral flow immunoassay test strips (optionally as an element of a chromatographic digital immunoassay) are well known in the art. For example, they typically contain an antibody specific to an antigen, and the specimen sample fluid migrates up the test strip and reacts with the antibody, thus generating a line on the test strip; the presence of this line indicates a positive test result. For example, the BD Veritor™ system for Rapid Detection of SARS-CoV-2 (Becton, Dickinson and Company, Maryland, USA, Ref: 256082) is a chromatographic digital immunoassay intended for the direct and qualitative detection of SARS-CoV-2 nucleocapsid antigens in nasal swabs.) Alternatively, for some applications, test 302 comprises a rapid molecular test kit, for example, an isothermal nucleic acid amplification (iNAAT) test, such as a rapid molecular test kit that uses a real-time loop mediated amplification reaction, such as the Lucira COVID-19 All-In-One Test Kit (Lucira Health, Inc., Emeryville, Calif., USA), or a nicking enzyme amplification reaction (NEAR) technology, such as the ID NOW™ (Abbott Laboratories, Abbott Park, Ill., USA), or a molecular test kit manufactured by Visby. Further alternatively, test 302 may comprise a CRISPR-based diagnostic test, an ELISA diagnostic test, or a spectroscopy-based diagnostic test.

[0609] Reference is now made to FIG. 13, which is a schematic illustration of a testing kit 400, in accordance with an application of the present invention. Testing kit 400 comprises nasal wash fluid system 100 (labeled in FIG. 2A), and further comprises a vial 402 and a liquid 404 for bathing at least a portion of specimen sample 24 in the vial, the liquid selected from the group consisting of: a lysis buffer, saline solution (e.g., phosphate buffered saline (PBS)), and transport medium (e.g., universal transport medium or a viral transport medium). Optionally, vial 402 contains liquid 404. Testing kit 400 may be used, for example, for collecting specimen sample 24 from the subject and sending at least a portion of the specimen sample to a remote laboratory in the liquid in the vial, such as for performing Polymerase Chain Reaction (PCR) testing for particulate in the specimen sample. (As described hereinabove, nasal wash fluid system 100 comprises nasal wash fluid dispenser 20 and nasal wash fluid 26.) The particulate may be a virus, a bacterium, any of the other particulates described hereinabove, or any of the other biological materials described hereinabove.

[0610] Reference is now made to FIGS. 14A-B, which are schematic illustrations of filter units 500A and 500B, respectively, for testing a liquid specimen sample that includes saliva for the presence of a particulate, in accordance with respective applications of the present invention. The particulate may be a virus, a bacterium, any of the other particulates described hereinabove, or any of the other biological materials described hereinabove.

[0611] Filter unit 500A comprises at least one filter chamber 510A (such as exactly one, as shown), which is shaped so as to define an inlet 512, an outlet 514, and a fluid flow path between inlet 512 and outlet 514. Filter unit 500B comprises at least one filter chamber 510B (such as exactly one, as shown), which is shaped so as to define inlet 512, outlet 514, and a fluid flow path between inlet 512 and outlet 514.

[0612] Each of the at least one filter chambers 510A and 510B comprises a large-pored filter 520, which is disposed within at the at least one filter chamber in the fluid flow path, and has a nominal pore size.

[0613] The at least one filter chamber 510B of filter unit 500A comprises large-pored filter 520, and one or more additional filters 522, which are disposed within the at the at least one filter chamber 510B in series in the fluid flow path, and which have respective nominal pore sizes, which may or may not equal one another, and may or may not equal the nominal pore size of large-pored filter 520. For example, the one or more additional filters 522 may include exactly one additional filter 522 (as shown) or more than one additional filter 522 (configuration not shown). Typically, but not necessarily, each of the respective nominal pore sizes of the one or more additional filters 522 is less than or equal to the nominal pore size of large-pored filter 520.

[0614] For some applications, the nominal pore size of large-pored filter 520 is between 30 microns and 1.5 mm.

[0615] For some applications, the nominal pore size is representative of a minimum size of spherical particles (e.g., inert spherical particles) necessary for the respective filter to retain 85% of the spherical particles when distilled H2O containing the spherical particles is passed through the respective filter at 20 degrees C. under pressure supplied by a 10 cm water column (of course, without downstream resistance) (herein, the “challenge-test nominal pore size”). It will be appreciated that the spherical particles, distilled H2O, and 10 cm water column are not elements of the device, but are instead non-device elements that are used to characterize certain physical properties of the filter(s), as is conventional for filter characterization in the filter art.

[0616] Alternatively, the nominal pore size may be characterized by a bubble point test, as is well known in the filter art, using a capillary flow porometer, such as CFP-1500A Automatic Capillary Flow Porometer (optionally, including options E, X, L, and/or M, if necessary for characterizing certain larger pore sizes) made by Porous Materials Inc. (PMI) (Ithaca, N.Y., USA), and a wetting liquid having a fluid surface tension of 15.9 dynes/cm, such as Galwick (PMI) (herein, the “bubble-point nominal pore size”).

[0617] As used in the present application, including in the claims and Inventive Concepts, a “pore” means any opening through a filter through which at least distilled H2O can pass.

[0618] For some applications, large-pored filter 520 comprises a depth filter.

[0619] For some applications, large-pored filter 520 comprises a mesh filter, such as a woven mesh filter or a non-woven mesh filter.

[0620] For some applications, large-pored filter 520 comprises fibers, such as crisscrossing fibers.

[0621] For some applications, large-pored filter 520 comprises a screen, such as a woven screen or a non-woven screen.

[0622] For some applications, large-pored filter 520 comprises a polymer, such as a polyester.

[0623] For some applications, large-pored filter 520 comprises a metal.

[0624] For some applications, the challenge-test nominal pore size is at least 40 microns. For some applications, the challenge-test nominal pore size is at least 50 microns. For some applications, the challenge-test nominal pore size is at least 60 microns. For some applications, the challenge-test nominal pore size is at least 100 microns. For some applications, the challenge-test nominal pore size is at least 120 microns. For some applications, the challenge-test nominal pore size is at least 150 microns. For some applications, the challenge-test nominal pore size is at least 200 microns. For some applications, the challenge-test nominal pore size is at least 500 microns.

[0625] For some applications, the challenge-test nominal pore size is less than 1 mm. For some applications, the challenge-test nominal pore size is less than 750 microns. For some applications, the challenge-test nominal pore size less than 500 microns. For some applications, the challenge-test nominal pore size less than 250 microns. For some applications, the challenge-test nominal pore size less than 200 microns.

[0626] For some applications, the challenge-test nominal pore size is between 30 and 40 microns. For some applications, the challenge-test nominal pore size is between 40 and 60 microns. For some applications, the challenge-test nominal pore size is between 50 and 200 microns, such as between 50 and 150 microns, e.g., between 100 and 150 microns. For some applications, the challenge-test nominal pore size is between 60 and 100 microns. For some applications, the challenge-test nominal pore size is between 100 and 120 microns. For some applications, the challenge-test nominal pore size is between 120 and 150 microns. For some applications, the challenge-test nominal pore size is between 150 and 200 microns. For some applications, the challenge-test nominal pore size is between 200 and 500 microns. For some applications, the challenge-test nominal pore size is between 500 microns and 1 mm, such as between 500 and 750 microns, or between 750 microns and 1 mm.

[0627] For some applications, the bubble-point nominal pore size is at least 40 microns. For some applications, the bubble-point nominal pore size is at least 50 microns. For some applications, the bubble-point nominal pore size is at least 60 microns. For some applications, the bubble-point nominal pore size is at least 100 microns. For some applications, the bubble-point nominal pore size is at least 120 microns. For some applications, the bubble-point nominal pore size is at least 150 microns. For some applications, the bubble-point nominal pore size is at least 200 microns.

[0628] For some applications, the bubble-point nominal pore size less than 500 microns. For some applications, the bubble-point nominal pore size less than 300 microns. For some applications, the bubble-point nominal pore size less than 250 microns. For some applications, the bubble-point nominal pore size less than 200 microns.

[0629] For some applications, the bubble-point nominal pore size is between 30 and 40 microns. For some applications, the bubble-point nominal pore size is between 40 and 60 microns. For some applications, the bubble-point nominal pore size is between 50 and 200 microns, such as between 50 and 150 microns, e.g., between 100 and 150 microns. For some applications, the bubble-point nominal pore size is between 60 and 100 microns. For some applications, the bubble-point nominal pore size is between 100 and 120 microns. For some applications, the bubble-point nominal pore size is between 120 and 150 microns. For some applications, the bubble-point nominal pore size is between 150 and 200 microns. For some applications, the bubble-point nominal pore size is between 200 and 300 microns.

[0630] For some applications, filter chambers 510A and 510B are generally cylindrical, as shown, while for other applications they may have different shapes.

[0631] For some applications, filter chambers 510A and 510B further comprise one or more filter supports 530, which are disposed within the filter chambers, and are shaped so as to define respective (a) proximal support surfaces that mechanically support respective filters, and (b) respective pluralities of filter-support openings through the filter supports to allow passage of filtrate from the respective filters.

[0632] For some applications, the filter unit further comprises a liquid-pressure source 534, which is arranged to apply pressure to drive the liquid specimen sample through the one or more filters. For example, liquid-pressure source 534 may comprise a pump 536, such as shown, by way of example, for filter unit 500A, or a plunger 538, such as shown, by way of example, for filter unit 500B.

[0633] For some applications, the filter unit further comprises a waste receptacle 540, which is disposed in fluid communication with outlet 514 of the filter chamber so as to receive filtrate from the one or more filters. For some of these applications, waste receptacle 540 cannot be removed from the filter chamber without damaging the filter unit, such as shown. For other applications, waste receptacle 540 is configured to be decoupled from the filter chamber without damaging the filter unit (configuration not shown).

[0634] For some applications, waste receptacle 540 contains a disinfectant 544, such as shown, by way of example, for filter unit 500A, or a liquid-absorbing material 546, such as shown, by way of example, for filter unit 500B.

[0635] In some applications of the present invention, a method is provided that comprises: [0636] collecting, from a subject, a liquid specimen sample that includes saliva and orally-dispensed oral wash fluid 68; [0637] passing the liquid specimen sample through a filter (such as large-pored filter 520, described hereinabove with reference to FIGS. 14A-B) to trap at least some of the saliva, the filter having a nominal pore size of between 30 microns and 1.5 mm; and [0638] testing for the presence of a virus in the saliva trapped (i.e., retained) by the filter.

[0639] Alternatively, the method comprises testing for the presence of a bacterium trapped by the filter. Further alternatively, the method comprises testing for the presence of another particulate trapped by the filter, such as any of the particulates mentioned hereinabove.

[0640] This method may be practiced in combination with any of the techniques described herein for collecting specimen sample 24 from the subject that include gargling by the subject, swishing by the subject, and/or spraying oropharynx 30 via anterior opening 34 of oral cavity 36.

[0641] As described below in the section entitled, “Experiment 7,” the inventors found that filters having pore sizes substantially larger than the viral diameter were nevertheless able to trap virus in the saliva of gargled fluid in quantities sufficient to clinically identify the presence of the virus, even using immunoassays, which generally require substantially greater quantities of virus for detection than does PCR. The inventors hypothesize that the large-pore filters were able to trap virus at least in part because of the viscosity of the saliva in typical gargled fluid samples, perhaps in particular because of the viscosity of the mucus and/or other large or aggregated salivary components of the saliva, which themselves contain viral particles, and/or because the viral particles adhered (e.g., directly adhered) to the material of the large-pore filters. Use of large-pore filters may enable a high flow rate and inhibit filter clogging despite high sample viscosity.

[0642] For some applications, the challenge-test nominal pore size is at least 40 microns. For some applications, the challenge-test nominal pore size is at least 50 microns. For some applications, the challenge-test nominal pore size is at least 60 microns. For some applications, the challenge-test nominal pore size is at least 100 microns. For some applications, the challenge-test nominal pore size is at least 120 microns. For some applications, the challenge-test nominal pore size is at least 150 microns. For some applications, the challenge-test nominal pore size is at least 200 microns. For some applications, the challenge-test nominal pore size is at least 500 microns.

[0643] For some applications, the challenge-test nominal pore size is less than 1 mm. For some applications, the challenge-test nominal pore size is less than 750 microns. For some applications, the challenge-test nominal pore size less than 500 microns. For some applications, the challenge-test nominal pore size less than 250 microns. For some applications, the challenge-test nominal pore size less than 200 microns.

[0644] For some applications, the challenge-test nominal pore size is between 30 and 40 microns. For some applications, the challenge-test nominal pore size is between 40 and 60 microns. For some applications, the challenge-test nominal pore size is between 50 and 200 microns, such as between 50 and 150 microns, e.g., between 100 and 150 microns. For some applications, the challenge-test nominal pore size is between 60 and 100 microns. For some applications, the challenge-test nominal pore size is between 100 and 120 microns. For some applications, the challenge-test nominal pore size is between 120 and 150 microns. For some applications, the challenge-test nominal pore size is between 150 and 200 microns. For some applications, the challenge-test nominal pore size is between 200 and 500 microns. For some applications, the challenge-test nominal pore size is between 500 microns and 1 mm, such as between 500 and 750 microns, or between 750 microns and 1 mm.

[0645] For some applications, the bubble-point nominal pore size is at least 40 microns. For some applications, the bubble-point nominal pore size is at least 50 microns. For some applications, the bubble-point nominal pore size is at least 60 microns. For some applications, the bubble-point nominal pore size is at least 100 microns. For some applications, the bubble-point nominal pore size is at least 120 microns. For some applications, the bubble-point nominal pore size is at least 150 microns. For some applications, the bubble-point nominal pore size is at least 200 microns.

[0646] For some applications, the bubble-point nominal pore size less than 500 microns. For some applications, the bubble-point nominal pore size less than 300 microns. For some applications, the bubble-point nominal pore size less than 250 microns. For some applications, the bubble-point nominal pore size less than 200 microns.

[0647] For some applications, the bubble-point nominal pore size is between 30 and 40 microns. For some applications, the bubble-point nominal pore size is between 40 and 60 microns. For some applications, the bubble-point nominal pore size is between 50 and 200 microns, such as between 50 and 150 microns, e.g., between 100 and 150 microns. For some applications, the bubble-point nominal pore size is between 60 and 100 microns. For some applications, the bubble-point nominal pore size is between 100 and 120 microns. For some applications, the bubble-point nominal pore size is between 120 and 150 microns. For some applications, the bubble-point nominal pore size is between 150 and 200 microns. For some applications, the bubble-point nominal pore size is between 200 and 300 microns.

[0648] Typically, the method does not comprise centrifuging the liquid specimen sample.

[0649] For some applications, the method does not comprise adding to the liquid specimen sample a concentration agent that binds to the virus.

[0650] For some applications, the filter is one of a plurality of filters arranged in a series of filters (such as described hereinabove with reference to FIG. 14B). For some applications, the filters have respective challenge-test nominal pore sizes, all of which are between 30 microns and 1.5 mm, or any of the challenge-test nominal pore sizes provided above. For some applications, the filters have respective bubble-point nominal pore sizes, all of which are between 30 and 500 microns, such as between 30 and 300 microns, or any of the bubble-point nominal pore sizes provided above. The method comprises passing the liquid specimen sample through the series of filters, without passing the liquid specimen sample through any other filters. Testing for the presence of the virus in the saliva comprises testing for the presence of the virus in the saliva trapped by at least one of the plurality of filters, such as testing for the presence of the virus in the saliva trapped by at least two of the plurality of filters.

[0651] For some applications, the filter is a first filter (such as large-pored filter 520 of filter chamber 510B, described hereinabove with reference to FIG. 14B). Passing the liquid specimen sample through the filter comprises passing the liquid specimen sample through the first filter to produce a filtrate. Testing for the presence of the virus in the saliva trapped by the filter comprises testing for the presence of the virus in the saliva trapped by the first filter. The method further comprises passing the filtrate through a second filter (such as additional filter 522 of filter chamber 510B, described hereinabove with reference to FIG. 14B) to trap at least some of the saliva, and testing for the presence of the virus in the saliva trapped by the second filter. For some of these applications, a nominal pore size of the second filter equals the nominal pore size of the first filter. For some of these applications, a challenge-test nominal pore size of the second filter is between 0.03 and 21 microns, such as between 0.8 and 1.5 microns. For some of these applications, a bubble-point nominal pore size of the second filter is between 0.03 and 21 microns, such as between 0.8 and 1.5 microns. For some of these applications, the second filter may be configured (such as by its nominal pore size) to capture epithelial cells.

[0652] For some applications, passing the liquid specimen sample through the filter comprises passing the liquid specimen sample through the filter to trap at least some of the saliva by size-based filtration.

[0653] For some applications, passing the liquid specimen sample through the filter comprises passing the liquid specimen sample through the filter to trap at least some of the saliva by adhesion of the saliva to the filter. For some applications, passing the liquid specimen sample through the filter to trap at least some of the saliva by adhesion of the saliva to the filter by interactions that are not specific to the virus (rather than specific interactions, such as ligand or antibody binding). For some applications, the filter is coated with organic molecules, such as lipids, proteins, and/or carbohydrates, to increase adhesion of the saliva to the filter.

[0654] For some applications, the filter comprises a depth filter.

[0655] For some applications, the filter comprises a mesh filter, such as a woven mesh filter or a non-woven mesh filter.

[0656] For some applications, the filter comprises fibers, such as crisscrossing fibers.

[0657] For some applications, the filter comprises a screen, such as a woven screen or a non-woven screen.

[0658] For some applications, the filter comprises a polymer, such as a polyester.

[0659] For some applications, the filter comprises a metal.

[0660] For some applications, passing the liquid specimen sample through the filter comprises passing the liquid specimen sample through exactly one filter.

[0661] For some applications, testing for the presence of the virus in the saliva trapped by the filter comprises testing the filter for the presence of the virus in the saliva.

[0662] For some applications, testing for the presence of the virus in the saliva trapped by the filter comprises inserting the filter into a testing machine. For example, the testing machine may be selected from the group consisting of: a thermal cycler and an isothermal amplification instrument.

[0663] For some applications, testing for the presence of the virus comprises performing a lateral flow immunoassay. As described below in the section entitled, “Experiment 7,” the inventors were able to detect the presence of SARS-CoV-2 in the biological material trapped by the filter using lateral flow immunoassays.

[0664] For some applications, testing for the presence of the virus in the saliva trapped by the filter comprises, after finishing passing the liquid specimen sample through the filter, bringing the filter, the saliva, and a liquid medium into contact, and testing the liquid medium for the presence of the virus. For some applications, the testing machine is selected from the group consisting of: a thermal cycler and an isothermal amplification instrument.

[0665] For some applications, testing the liquid medium for the presence of the virus comprises separating the liquid medium from the filter before testing the liquid medium for the presence of the virus. For some applications, testing for the presence of the virus trapped by the filter comprises agitating the filter in the liquid medium, for example mixing the filter in the liquid medium.

[0666] For some applications, testing for the presence of the virus trapped by the filter comprises transferring at least a portion of the liquid medium into a testing machine.

[0667] For some applications, the liquid medium is selected from the group consisting of: a transport medium, a culture medium, a purification agent, a stabilizing agent, a lysing buffer, and an extraction agent.

[0668] For some applications, testing for the presence of the virus comprises using spectroscopy, using microscopy, using image processing, and/or using three-dimensional (3D) image sensing. For some applications, testing for the presence of the virus comprises using a molecular-based assay. For some of these applications, testing for the presence of the virus comprises using nucleic acid amplification, such as isothermal amplification or polymerase chain reaction (PCR). As described below in the section entitled, “Experiment 7,” the inventors were able to detect the presence of SARS-CoV-2 in the biological material trapped by the filter using PCR.

[0669] For some applications, testing for the presence of the virus comprises using artificial intelligence. Any of the above-mentioned testing techniques may optionally utilize artificial intelligence.

[0670] For some applications, the method further comprises, before testing for the presence of the virus in the saliva trapped by the filter, transporting the filter while the filter is at least partially immersed in a liquid medium. For some of these applications, testing for the presence of the virus in the saliva trapped by the filter comprises inserting at least a portion of the liquid medium into a thermal cycler. For some of these applications, the filter is placed in the liquid medium, the liquid medium is agitated, and the filter is removed from the liquid medium before transporting the liquid medium to a remote testing site for testing for the presence of the virus in the saliva trapped by the filter.

[0671] For some applications, collecting the liquid specimen sample comprises receiving the liquid specimen sample by a healthcare worker from the subject.

[0672] For some applications, the method does not comprise swabbing nasal cavity 22. Alternatively or additionally, for some applications, the method does not comprise swabbing a pharynx of the subject.

[0673] For some applications, the method does not comprise swabbing any portion of the subject. Alternatively, the method comprises swabbing a portion of the subject, such as using swabbing techniques described herein.

[0674] For some applications, collecting the liquid specimen sample comprises collecting the liquid specimen sample that was expressed out of oropharynx 30 via anterior opening 34 of oral cavity 36. For some of these applications, collecting the liquid specimen sample comprises collecting the liquid specimen sample that was spit out of oropharynx 30 via anterior opening 34 of oral cavity 36.

[0675] For some applications, collecting the liquid specimen sample comprises aspirating the liquid specimen sample out of oropharynx 30 via anterior opening 34 of oral cavity 36.

[0676] For some applications, collecting the liquid specimen sample comprises: [0677] gargling, by the subject, oral wash fluid 68 to produce the liquid specimen sample that includes the saliva; and [0678] thereafter, collecting the liquid specimen sample that passed out of anterior opening 34 of oral cavity 36.

[0679] For some applications, collecting the liquid specimen sample comprises: [0680] gargling, by the subject, oral wash fluid 68 to produce the liquid specimen sample that includes the saliva; and [0681] thereafter, expressing, by the subject, out of anterior opening 34 of oral cavity 36, the liquid specimen sample.

[0682] For some applications, collecting the liquid specimen sample comprises: [0683] swishing, by the subject, oral wash fluid 68 in oral cavity 36 to produce the liquid specimen sample that includes the oral wash fluid and the saliva; and [0684] thereafter, collecting the liquid specimen sample from the subject.

[0685] For some applications, collecting the liquid specimen sample comprises: [0686] swishing, by the subject, oral wash fluid 68 in oral cavity 36 to produce the liquid specimen sample that includes the oral wash fluid and the saliva; and [0687] thereafter, expressing, by the subject, out of anterior opening 34 of oral cavity 36, the liquid specimen sample.

[0688] For some applications, collecting the liquid specimen sample comprises: [0689] spraying, via anterior opening 34 of oral cavity 36, oropharynx 30 with oral wash fluid 68; and [0690] thereafter, collecting the liquid specimen sample from the subject.

[0691] For some applications, the above-mentioned gargling, swishing, and/or spraying techniques are combined.

[0692] For some applications, collecting the gargled fluid from the subject comprises: [0693] intranasally dispensing nasal wash fluid 26 into nasal cavity 22 such that nasal wash fluid 26 washes biological material into oropharynx 30 from (a) nasal cavity 22, (b) nasopharynx 32, or (c) nasal cavity 22 and nasopharynx 32; and [0694] thereafter, collecting the liquid specimen sample that passed out of anterior opening 34 of oral cavity 36 and includes oral wash fluid 68, nasal wash fluid 26, and at least a portion of the biological material washed into oropharynx 30 by nasal wash fluid 26.

[0695] For some applications, collecting the liquid specimen sample comprises expressing, by the subject, the liquid specimen sample out of oropharynx 30 via anterior opening 34 of oral cavity 36. For some applications, expressing the liquid specimen sample comprises spitting, by the subject, the liquid specimen sample out of oropharynx 30 via anterior opening 34 of oral cavity 36.

[0696] For some applications, collecting the liquid specimen sample comprises: [0697] after intranasally dispensing nasal wash fluid 26, orally dispensing oral wash fluid into oral cavity 36 via anterior opening 34 of oral cavity 36 such that oral wash fluid 68 forms mixture 70 with nasal wash fluid 26 in oropharynx 30; [0698] gargling, by the subject, mixture 70 to produce the liquid specimen samples that includes oral wash fluid 68, nasal wash fluid 26, and the saliva; and [0699] collecting the liquid specimen sample that passed out of anterior opening 34 of oral cavity 36.

[0700] For some applications, collecting the liquid specimen sample from the subject comprises: [0701] intranasally dispensing nasal wash fluid into nasal cavity 22 such that nasal wash fluid 26 washes biological material into an oropharynx of the subject from (a) nasal cavity 22, (b) a nasopharynx of the subject, or (c) nasal cavity 22 and the nasopharynx; and [0702] thereafter, expressing, by the subject, out of anterior opening 34 of oral cavity 36, the liquid specimen sample that includes at least a portion of the biological material washed into oropharynx 30 by nasal wash fluid 26.

[0703] For some applications, expressing the liquid specimen sample comprises spitting, by the subject, the liquid specimen sample out of oropharynx 30 via anterior opening 34 of oral cavity 36.

[0704] For some applications, collecting the liquid specimen sample comprises: [0705] after intranasally dispensing nasal wash fluid 26, orally dispensing oral wash fluid 68 into oral cavity 36 via anterior opening 34 of oral cavity 36 such that oral wash fluid 68 forms mixture 70 with nasal wash fluid 26 in oropharynx 30; [0706] gargling, by the subject, mixture 70 to produce the liquid specimen sample that includes oral wash fluid 68, nasal wash fluid 26, and the saliva; and [0707] expressing, by the subject, the liquid specimen sample out of anterior opening 34 of oral cavity 36.

[0708] For some applications, the subject intranasally dispenses nasal wash fluid 26 into nasal cavity 22, while for some other applications, a healthcare worker intranasally dispenses nasal wash fluid 26 into nasal cavity 22.

[0709] For some applications, the method further comprises, after intranasally dispensing nasal wash fluid 26, gargling, by the subject, nasal wash fluid 26 washed into oropharynx 30.

[0710] For some applications, the virus is a coronavirus, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2); or an Influenza virus.

[0711] For some applications, the subject is one of a plurality of subjects, and wherein the liquid specimen sample is one of a plurality of liquid specimen samples. The plurality of liquid specimen samples, which include respective saliva, are collected from the plurality of subjects, respectively. The plurality of liquid specimen samples are combined, and the combined plurality of liquid specimen samples are passed through the filter. For example, the plurality of liquid specimen samples may include at least five liquid specimen samples from at least five subjects, respectively, such as at least ten liquid specimen samples from at least ten subjects, respectively, or at least 25 liquid specimen samples from at least 25 subjects, respectively.

[0712] This method of combining liquid specimen samples from a plurality of subjects allows screening a plurality of subjects using a single filter unit and a single test of the material captured by the one or more filters of the single filter unit. If the particulate, e.g., the virus, is detected in the combined sample, the subjects may be individually tested.

[0713] For some applications, the plurality of liquid specimen samples have a combined volume of at least 25 cc, such as at least 50 cc, e.g., at least 100 cc, or at least 150 cc.

[0714] For some applications, combining the plurality of liquid specimen samples comprises mixing together the plurality of liquid specimen samples.

[0715] Any of the method described herein that comprise collecting, filtering, and testing a liquid specimen sample that includes saliva and orally-dispensed oral wash fluid 68 may alternatively comprise collecting, filtering, and testing a liquid specimen sample that includes (a) a biological material other than saliva and (b) orally-dispensed oral wash fluid 68.

[0716] Reference is now made to FIG. 15, which is a schematic illustration of a testing kit 600, in accordance with an application of the present invention. Testing kit 600 comprises a filter unit 610, such as one of filter units 500A or 500B, described hereinabove with reference to FIGS. 14A-B, respectively; and a test 602 (e.g., a diagnostic test), which is configured to test for the presence of a particulate in the virus or other particulate (such as those described herein) trapped by the one or more filters of the filter unit. Test 602 may optionally be configured to implement testing techniques described herein, in the patent applications incorporated hereinbelow by reference, and/or known in the art. For some applications, test 602 comprises a lateral flow immunoassay test strip, which is configured to detect the presence of the particulate (such as by detecting protein antigen, e.g., from a virus), and, optionally, one or more reagents. Alternatively, for some applications, test 602 comprises a rapid molecular test kit, for example one that uses a real-time loop mediated amplification reaction, such as the Lucira COVID-19 All-In-One Test Kit, or a NEAR technology, such as the ID NOW™, or a molecular test kit manufactured by Visby. Further alternatively, test 302 may comprise a CRISPR-based diagnostic test, an ELISA diagnostic test, or a spectroscopy-based diagnostic test.

[0717] Reference is now made to FIG. 16, which is a schematic illustration of a testing kit 700, in accordance with an application of the present invention. Testing kit 700 comprises: [0718] filter unit 610, such as one of filter units 500A or 500B, described hereinabove with reference to FIGS. 14A-B, respectively; [0719] a vial 702, for containing at least a portion of the saliva captured by at least one of the one or more filters of the filter unit; and [0720] liquid 704 for bathing the at least a portion of the saliva in the vial, the liquid selected from the group consisting of: a lysis buffer, saline solution (e.g., phosphate buffered saline (PBS)), and transport medium (e.g., universal transport medium or a viral transport medium).

[0721] Optionally, vial 702 contains liquid 704. Testing kit 700 may be used, for example, for collecting a specimen sample from the subject and sending at least a portion of the specimen sample to a remote laboratory in the liquid in the vial, such as for performing Polymerase Chain Reaction (PCR) testing for particulate in the specimen sample. The particulate may be a virus (e.g., an Influenza virus or SARS-CoV-2), a bacterium (e.g., Streptococcus bacterium), any of the other particulates described hereinabove, or any of the other biological materials described hereinabove. For example, the particulate may be tested for using nucleic acid amplification, such as PCR, e.g., qPCR, and/or by performing an immunoassay, such as a lateral flow immunoassay, e.g., a chromatographic digital immunoassay, or by performing a rapid molecular test, for example one that uses a real-time loop mediated amplification reaction, such as the Lucira COVID-19 All-In-One Test Kit, or a NEAR technology, such as the ID NOW™, or a molecular test kit manufactured by Visby. Further alternatively, test 302 may comprise a CRISPR-based diagnostic test, an ELISA diagnostic test, or a spectroscopy-based diagnostic test.

[0722] Reference is made to both FIGS. 15 and 16. For some applications, testing kit 600 or 700 does not comprise any specimen-collection swabs.

[0723] For some applications, testing kit 600 or 700 comprises a nasal applicator containing nasal wash fluid 26, such as a conventional nasal dispenser or a nasal wash fluid dispenser described hereinabove. For some of these applications, the testing kit comprises a plurality of nasal applicators containing nasal wash fluid 26.

[0724] For some applications, testing kit 600 or 700 comprises a first number of nasal applicators, and a second number of one or more filter units 610, and a ratio of the first number to the second number is at least 2, such as at least 5.

[0725] For some applications, testing kit 600 or 700 further comprises a container containing oral wash fluid 68. For some applications, the testing kit comprises a plurality of nasal applicators containing nasal wash fluid 26 and a plurality of containers containing oral wash fluid 68, and the testing kit comprises a first number of nasal applicators containing nasal wash fluid 26, a second number of one or more filter units 610, and a third number of containers containing oral wash fluid 68. The first number equals the third number, and a ratio of the first number to the second number is at least 2, such as at least 5.

[0726] For some applications, testing kit 600 or 700 further comprises a container containing oral wash fluid 68. For some of these applications, the test kit comprises a plurality of containers containing oral wash fluid 68. For some of these applications, the testing kit comprises a third number of containers containing oral wash fluid 68, and a second number of one or more filter units 610, and a ratio of the third number to the second number is at least 2, such as at least 5.

[0727] Reference is now made to FIGS. 17A-B, which are schematic illustrations of a method for collecting a specimen sample from a subject, in accordance with an application of the present invention. The method comprises intranasally dispensing nasal wash fluid 26 into nasal cavity 22, by the subject or by somebody else. Nasal wash fluid 26 may be intranasally dispensed using a conventional nasal dispenser 820, such as shown in FIG. 17A; using nasal wash fluid dispenser 20, such as shown in FIG. 2B; using nasal sampling device 1020, such as shown in FIG. 22; or using any nasal wash fluid dispenser known in the art. Nasal wash fluid 26 generally loosens biological material in nasal cavity 22 and/or nasopharynx 32. Thereafter, a specimen sample is collected by performing, by the subject or by somebody else, one or more nasal swabs selected from the group consisting of: an anterior nares swab (such as shown in FIG. 17B) and a mid-turbinate swab (not shown). The nasal swab may be performed using a conventional nasal swab 822, such as shown in FIG. 17B, or using nasal sampling device 1020, such as shown in FIG. 22. The specimen sample is typically collected without performing a nasopharyngeal swab. When the specimen sample is collected by performing the anterior nares swab, the specimen sample is optionally collected without performing a mid-turbinate swab or a nasopharyngeal swab. The nasal swab may be performed in one or both of the nostrils; in the latter case, nasal wash fluid 26 is intranasally dispensed into both nostrils of nasal cavity 22, and the same single swab is typically used in both nostrils. Alternatively or additionally, the specimen sample is collected by performing a nasopharyngeal swab.

[0728] In some applications of the present invention, a method is provided that comprises intranasally dispensing nasal wash fluid 26 into nasal cavity 22 of a subject, by the subject or somebody else, such as shown in FIG. 17A. Nasal wash fluid 26 generally loosens biological material in nasal cavity 22 and/or nasopharynx 32. Thereafter, a specimen sampler is inserted into nasal cavity 22 by the subject or by somebody else, such that a farthest-inserted portion of the specimen sampler is positioned in an anterior naris of nasal cavity 22. A specimen sample is collected on the specimen sampler by contacting (e.g., rubbing) a wall of the anterior naris (i.e., the wall's epithelial surface) with the specimen sampler, optionally including rotating the specimen sampler. For example, the specimen sampler may comprise a swab (e.g., a flocked swab) (such as shown in FIG. 17B), a brush, a sponge, or a nasal plug. Typically, but not necessarily, the specimen sampler comprises a tip comprising an absorbent material. For applications in which the specimen sampler comprises a swab, a tip of the swab may, for example, be a flocked swab tip (e.g., comprising nylon), a rayon swab tip, a cotton swab tip, a polyurethane foam swab tip, a polyethylene terephthalate (PET) swab tip, a polyester swab tip, a foam swab tip (e.g., comprising polyurethane) (e.g., an open-cell foam swab), a sponge swab tip (e.g., comprising cellulose), a brush swab tip (e.g., similar to a cytology brush), and a nanofiber swab tip. The specimen sampler may be inserted into a single nostril of nasal cavity 22, or into both of the nostrils, one at a time; in the latter case, nasal wash fluid 26 is intranasally dispensed into both nostrils of nasal cavity 22.

[0729] In some applications of the present invention, a method is provided that comprises intranasally dispensing nasal wash fluid 26 into nasal cavity 22 of a subject, by the subject or somebody else, such as shown in FIG. 17A. Nasal wash fluid 26 generally loosens biological material in nasal cavity 22 and/or nasopharynx 32. Thereafter, a specimen sampler is inserted into nasal cavity 22 by the subject or by somebody else. While the specimen sample is inserted into the nasal cavity, a specimen sample is collected on the specimen sampler without using suction, such as shown in FIG. 17B.

[0730] In some applications of the present invention, a method is provided that comprises intranasally dispensing nasal wash fluid 26 into nasal cavity 22 of a subject, by the subject or somebody else, such as shown in FIG. 17A. Nasal wash fluid 26 generally loosens biological material in nasal cavity 22 and/or nasopharynx 32. Thereafter, the subject or somebody else inserts, into nasal cavity 22, a specimen sampler that is shaped so as not to define any dispensing openings. While the specimen sampler is inserted into the nasal cavity, a specimen sample is collected on the specimen sampler, such as shown in FIG. 17B.

[0731] For some applications, before the specimen sample is collected using one of the techniques described in the previous paragraphs (by performing the nasal swab or using a specimen sampler inserted into the nasal cavity): [0732] before nasal wash fluid 26 is intranasally dispensed, the subject (typically while sitting or standing) tilts back his or her head (typically until the neck is fully extended) and holds the head tilted back while nasal wash fluid 26 is intranasally dispensed, or [0733] after nasal wash fluid 26 is intranasally dispensed, the subject tilts his or her head farther back than while intranasally dispensing the nasal wash fluid and farther back than a neutral position.

[0734] In all of these cases, the tilting back may cause nasal wash fluid 26 to loosen biological material in a more posterior (deeper) portion of nasal cavity 22 and/or nasopharynx 32.

[0735] Optionally, the subject sniffs after nasal wash fluid 26 is intranasally dispensed and before the specimen sample is collected. In applications in which the subject tilts the head far back, the sniffing may be performed before or after such far back tilting.

[0736] For some applications, nasal wash fluid 26 is intranasally dispensed without blowing, by the subject, a nose of the subject immediately prior to intranasally dispensing the nasal wash fluid. Blowing the nose may undesirably remove mucus and other biological material containing high concentrations of particulate of interest. Alternatively, the subject may blow his or her nose before the nasal wash fluid is intranasally dispensed.

[0737] Optionally, after the subject tilts his or her head back and after nasal wash fluid 26 is intranasally dispensed, the subject tilts forward his or her head to a neutral or a tilted-forward position; this backward-followed-by-forward tilting may help move biological material anteriorly (i.e., toward the nostril) within nasal cavity 22.

[0738] Optionally, the method further comprises, while holding his or her head tilted back, waiting for a period of time between (a) concluding intranasally dispensing nasal wash fluid 26 into the nasal cavity or tilting the head farther back and (b) tilting his or her head forward to the neutral or the tilted-forward position, the period of time at least 2 seconds (e.g., at least 5 seconds, such as at least 10 seconds, e.g., at least 15 seconds), no more than 60 seconds (e.g., no more than 30 seconds), and/or between 2 seconds (e.g., 5 seconds, such as 10 seconds, e.g., at least 15 seconds) and 60 seconds (e.g., 30 seconds).

[0739] Optionally, while holding his or her head tilted back, the subject makes a sound, such as a “K” sound, that inhibits flow of nasal wash fluid 26 from nasopharynx 32 to oropharynx 30, helping retain nasal wash fluid 26 in nasopharynx 32.

[0740] For some applications, the subject tilts his or her head forward to the neutral or the tilted-forward position after the subject senses (e.g., by feeling and/or tasting) the nasal wash fluid in oropharynx 30. Optionally, nasal wash fluid 26 comprises a tastant, for example, menthol flavoring.

[0741] For some applications, after tilting the head forward to the neutral or the tilted-forward position, the subject waits for a period of time before collecting the specimen sample, the period of time at least 5 seconds, e.g., at least 10 seconds, such as at least 30 seconds (and typically less than 120 seconds). This waiting period may allow sufficient time for nasal wash fluid 26 to drip anteriorly in nasal cavity 22, bringing the biological material with it.

[0742] Alternatively, for some applications, nasal wash fluid 26 is intranasally dispensed into nasal cavity 22 (either continuously or in a plurality of pulses) until the subject senses (e.g., by feeling and/or tasting) nasal wash fluid 26 in oropharynx 30. Optionally, nasal wash fluid 26 comprises a tastant, for example, menthol flavoring.

[0743] Optionally, the method further comprises waiting for a period of time between (a) concluding intranasally dispensing nasal wash fluid 26 into the nasal cavity and (b) collecting the specimen sample, the period of time at least 2 seconds (e.g., at least 5 seconds, such as at least 10 seconds), no more than 30 minutes (e.g., no more than 15 minutes), and/or between 2 seconds (e.g., 5 seconds, such as at least 10 seconds) and 30 minutes (e.g., 15 minutes).

[0744] For some applications, nasal wash fluid 26 is intranasally dispensed as one or more types of fluid discharge selected from the group consisting of: a spray, a mist, a pressurized aerosol, steam, and an atomized fluid. For some applications, nasal wash fluid 26 is intranasally dispensed as a fluid stream.

[0745] For some applications, nasal wash fluid 26 is dispensed into each of one or both nostrils 28 in a total volume of between 0.15 and 1 mL, such as between 0.15 and 0.5 mL, or between 0.25 and 1 mL, such as between 0.25 and 0.5 mL. Optionally, nasal wash fluid 26 is intranasally dispensed in a plurality of pulses per nostril (e.g., between 3 and 5 pulses per nostril), each having a volume of between 0.05 and 0.2 mL, for example, between 0.05 and 0.15 mL, e.g., 0.1 mL. (If nasal wash fluid 26 is dispensed into both nostrils 28, the volumes above would be doubled.) Dispensing such relatively small volumes reduces the likelihood of nasal wash fluid 26 washing out the biological material of interest from the nostrils.

[0746] In some applications, the specimen samples collected using the collection techniques described herein are tested for a particulate using one or more of the techniques described herein, one or more of the techniques described in the patent applications incorporated hereinbelow by reference, or a technique known in the art. The particulate may be a virus (e.g., SARS-CoV-2 or an Influenza virus), a bacterium, any of the other particulates described hereinabove, or any of the other biological materials described hereinabove. For example, the particulate may be tested for the presence of a particulate using nucleic acid amplification, such as PCR, e.g., qPCR, and/or by performing an immunoassay, such as a lateral flow immunoassay, e.g., a chromatographic digital immunoassay, or by performing a rapid molecular test, for example one that uses a real-time loop mediated amplification reaction, such as the Lucira COVID-19 All-In-One Test Kit, or a NEAR technology, such as the ID NOW™, or a molecular test kit manufactured by Visby. Further alternatively, the particulate may be tested the presence of a particulate using a CRISPR-based diagnostic test, an ELISA diagnostic test, or a spectroscopy-based diagnostic test.

[0747] These sampling methods may be combined with any other applicable techniques described herein, mutatis mutandis. By way of example and not limitation, these sampling methods may further comprise, after nasal wash fluid 26 is intranasally dispensed, orally dispensing oral wash fluid 68 into oral cavity 36 via anterior opening 34 of oral cavity 36; and gargling, by the subject, oral wash fluid 68. Optionally: [0748] oral wash fluid 68 is orally dispensed into oral cavity 36 such that oral wash fluid 68 forms mixture 70 with nasal wash fluid 26 in oropharynx 30, and, in addition to collecting the specimen sample from the nasal cavity (by swabbing or using the specimen sampler), a specimen sample 24 is collected that passed out of anterior opening 34 of oral cavity 36 and contains at least a portion of mixture 70 and, typically, at least a portion of biological material washed into oropharynx 30 with nasal wash fluid 26; and/or [0749] oral wash fluid 68 is orally dispensed into oral cavity 36, the subject gargles oral wash fluid 68, such that oral wash fluid 68 forms mixture 70 with nasal wash fluid 26, and a specimen sample 24 is collected that passed out of anterior opening 34 of oral cavity 36 and contains at least a portion of mixture 70 and the at least a portion of biological material washed into oropharynx 30 with nasal wash fluid 26.

[0750] Alternatively, a specimen sample is not collected that passed out of anterior opening 34 of oral cavity 36, and the gargled fluid is instead spit out and discarded. In this case, the time spent gargling (e.g., 5-15 seconds) may serve to ensure that the subject holds his or her head tilted back for a sufficient period of time for nasal wash fluid 26 to loosen biological material in nasal cavity 22 (such as in a more posterior (deeper) portion of nasal cavity 22 and/or nasopharynx 32). The gargling may alternatively or additionally distract the subject from the unpleasant sensation of post-nasal drip of nasal wash fluid 26 into the throat, so that the subject does not prematurely tilt his or her head forward to the neutral or the tilted-forward position before nasal wash fluid 26 sufficiently loosens the biological material in nasal cavity 22.

[0751] Also by way of example and not limitation, these nasal sampling methods may be performed using any of nasal wash fluid 26 dispensers described herein with reference to FIGS. 1, 2A-C, 3A-B, 6, 7A-K, 8A-C, 9A-F, 10, and/or 11. Alternatively, these nasal sampling methods may be performed using a conventional nasal dispenser, such as shown in FIG. 17A. Further alternatively, these nasal sampling methods may be performed using nasal sampling device 1020, described hereinbelow with reference to FIGS. 20A-B and 21A-B.

[0752] Experiments 9 and 10, described hereinbelow, provide evidence of the efficacy of some of these nasal-spray and nasal-sampling techniques for collecting specimen samples for detection of SARS-CoV-2 using quantitative PCR (qPCR).

[0753] Experiments 11 and 13, described hereinbelow, provide evidence of the efficacy of some of these nasal-spray and nasal-sampling techniques for collecting specimen samples for detection of SARS-CoV-2 using lateral flow immunoassay strips. The inventors believe that one of ordinary skill in the art would not expect that intranasal dispensation of nasal wash fluid 26 would enhance the sensitivity of detection of SARS-CoV-2 (or any other virus or biological particulate) using a lateral flow immunoassay strip, because of the much lower sensitivity of lateral flow immunoassay strips than nucleic acid amplification tests, such as PCR. Lateral flow immunoassay strips are well known to provide accurate results only when a fairly dry and concentrated sample is immersed in a small volume of reagent liquid, such as 0.2 to 0.4 mL, before application to the strip. In addition, anterior nares samples are known to be weaker than mid-turbinate and nasopharyngeal samples. Therefore, one of ordinary skill in the art would expect that the intranasal dispensation of nasal wash fluid 26 might dilute the biological material captured in the anterior nares specimen sample. In addition, one of ordinary skill in the art would expect that the intranasal dispensation of nasal wash fluid 26 might wash biological material potentially containing the virus off of the wall of the anterior nares before collection of the specimen sample, thereby reducing the amount of virus collected from the anterior nares.

[0754] For all of these reasons, one of ordinary skill in the art would expect the intranasal dispensation of nasal wash fluid before collecting the specimen sample would reduce the sensitivity of tests performed using lateral flow immunoassay strips. By contrast, the inventors have found that intranasal dispensation of nasal wash fluid 26 improves the sensitivity of lateral flow immunoassay tests performed on nasal samples collected from the anterior nares.

[0755] The dimensions and absorptive properties of swabs that may be useful in the nasal swabbing sample collection techniques described herein, such as for testing using a lateral flow immunoassay strip (e.g., for SARS-CoV-2 or an Influenza virus) were examined on behalf of the inventors. Specifically, flocked nasal swabs (3BY Ltd., Industrial Park Tefen, Israel, Ref: 3BY-001) were examined, because they were used in a number of the experiments described herein, and foam tipped nasal swabs (Puritan, Me., USA, Ref: 25-1506-1PF 100), which were provided with the BD Veritor™ system for Rapid Detection of SARS-CoV-2 (Becton Dickinson and Company, Maryland, USA, Ref: 256082), were examined because they are appropriate for sample collection for use in immunoassays.

[0756] Swab tip dimensions were determined using a mechanical caliper, or were provided by swab manufacturers (Table 1). Following saline nasal spraying and swabbing, swabs were found to contain between approximately 76-131 microliters of fluid from a single nostril (Table 2), after administering about 500 microliters of nasal wash into the nostril. Utilizing relatively smaller swab tips for respiratory specimen collection may be beneficial because of the limited volumes of lysing buffer included in commercial immunoassay kits (Table 3), which must be sufficient for the extraction of viral antigens from virions, intact infected cells, or infected cell debris which have been collected by the swab. If swabs were to contain a larger sample volume, the lysing buffer would be diluted beyond the optimal threshold and effective extraction would be unlikely to occur. Moreover, oversized swabs which absorb more fluid than what is recoverable during sampling would further absorb lysing buffer, making it difficult to elute sufficient lysate from the swab for assaying, because standard immunoassay cassettes require 3 drops of lysate (approximately 150 microliters), with dipstick style immunoassays typically absorbing at least 20 microliters of lysate.

TABLE-US-00001 TABLE 1 Swab Dimensions Swab Type Flocked Foam Greatest Tip Diameter (mm).sup.v 4 4.775.sup.i Tip Length (mm) 17 16.002.sup.i Tip Surface Area (mm.sup.2).sup.ii 226 258 Tip Volume (mm.sup.3).sup.iii 214 286 Applicator Thickness at Tip (mm) 2 2.54.sup.i Applicator Length at Tip (mm) 16.5 13 Applicator Volume at Tip (mm2) .sup.iii 52 64 Effective Absorptive Volume (mm.sup.3).sup.iii,iv 162 222 .sup.iDimensions provided by manufacturer (Puritan Medical Products Company LLC, Guilford, Maine, USA) .sup.iiSurface area was calculated assuming a perfectly cylindrical shape but excluding the bottom of the tip because of the presence of the applicator (2πrh + πr{circumflex over ( )}2). .sup.iiiVolume was calculated assuming a perfectly cylindrical shape. .sup.ivEffective absorptive volume was determined by subtracting the applicator volume at tip from the tip volume. .sup.ivThe greatest tip diameter is the diameter of the swab at the widest location along the swab.

TABLE-US-00002 TABLE 2 Swab Absorption Weight Weight Before After Fluid Swabbing Swabbing Swabbing Difference Volume Type Swab (g) (g) (g) (microliters).sup.i Saturation with Flocked 0.668 0.768 0.1 100 DPBS Foam 0.468 0.502 0.034 34 (−Rotation).sup.ii Saturation with Flocked 0.691 0.795 0.104 104 DPBS Foam 0.476 0.651 0.175 175 (+Rotation).sup.ii Bilateral Flocked 0.669 0.672 0.003 3 Swabbing Foam 0.497 0.493 −0.004 −4.sup.iv Spraying and Flocked 0.672 0.768 0.096 96 Unilateral Foam 0.472 0.548 0.076 76 Swabbing.sup.iii Spraying and Flocked 0.678 0.783 0.105 105 Bilateral Foam 0.469 0.6 0.131 131 Swabbing.sup.iii .sup.iAssumed density of 1 g/cm3 for absorbed liquid. .sup.iiSwabs were dipped into a 50 mL conical tube containing Deulbecco's Phosphate Buffered Saline (DBPS), and them either removed (−rotation) or rotated while brushing the walls of the tube five times to simulate anterior nares swabbing (+rotation) .sup.iiiUnilateral and Bilateral swabbing of the anterior nares were performed by inserting the entire tip of the swab into the nares and rotating the swab around the inner walls of the nares. Saline nasal spraying consisted of 5 sprays of 0.1 mL of saline into one or both nostriles. .sup.ivNegative values are assumed to be a result of either a small amount of the tip separating from the swab during swabbing, or are an artifcat due to inaccuracies in the analytical balance.

TABLE-US-00003 TABLE 3 Lysing Buffer Volumes Weight of Approximate Lysing Volume Assay Name Manufacturer Buffer (g) (microliters).sup.i QuickVue Influenza Quidel Corporation 0.245 245 A + B Test (San Diego, CA, Ref: 20183) BD Veritor ™ Becton Dickinson 0.413 413 system for Rapid and Company Detection of SARS- (Maryland, USA, CoV-2 Ref: 256082) COVID-19 Antigen Beijing Kewei 0.283 283 Rapid Test Cassette Clinical Diagnostic Reagent Inc (Beijing, China, Ref: 601450) .sup.iAssumed density of 1 g/cm3 for liquid reagents

[0757] For some applications, any of the swabs used in the methods described herein, or as elements of apparatus described herein, comprise a tip having one or more of the following characteristics: [0758] a greatest diameter of at least 2 mm, less than 5 mm (e.g., less than 4.5 mm), and/or between 2 and 5 mm (e.g., between 2 and 4.5 mm), [0759] a collection surface area of at least 200 mm2, less than 300 mm2, and/or between 200 and 300 mm2 (the collection surface area includes the absorptive portion of the swab, but excludes the bottom of the tip because of the presence of the applicator), [0760] a volume of at least 200 mm3, less than 300 mm3, and/or between 200 and 300 mm3, and/or [0761] a length of at least 12 mm, less than 20 mm, and/or between 12 and 20 mm.

[0762] Optionally, any of the swabs and/or specimen samplers described herein, including, but not limited to, with reference to FIGS. 18, 19, 20A-B, 21A-B, 21C-D, 22, and 23A-B, may have the characteristics described immediately above.

[0763] For some applications, in any of the nasal collection techniques described herein, collecting the specimen sample comprises collecting at least 25 microliters, less than 150 microliters (e.g., less than 125 microliters), and/or between 25 and 150 microliters (e.g., between 25 and 125 microliters) of material per specimen sampler, e.g., per swab (such as, for example, flocked swab). The material typically includes a combination of the biological material of interest and a portion of nasal wash fluid 26 dispensed into nasal cavity 22. Alternatively, for some applications, in any of the nasal collection techniques described herein, collecting the specimen sample comprises collecting at least 25 microliters, less than 250 microliters (e.g., less than 200 microliters), and/or between 25 and 250 microliters (e.g., between 25 and 200 microliters) of material per specimen sampler, e.g., per swab (such as, for example, foam swab).

[0764] In some applications of the present invention, a method is provided that comprises intranasally dispensing nasal wash fluid 26 into nasal cavity 22 of a subject, by the subject or by somebody else. Nasal wash fluid 26 generally loosens biological material in nasal cavity 22 and/or nasopharynx 32. Thereafter, a specimen sample is collected by blowing a nose, by the subject, into a specimen receptacle. These techniques may be practiced in combination with any of the techniques described herein for intranasally dispensing nasal wash fluid 26 into nasal cavity 22.

[0765] Reference is now made to FIG. 18, which is a schematic illustration of a testing kit 800, in accordance with an application of the present invention. Testing kit 800 comprises: [0766] a nasal dispenser (containing nasal wash fluid 26), such as conventional nasal dispenser 820, as shown, or nasal wash fluid dispenser 20, not shown in FIG. 18, but shown in FIG. 2B; [0767] conventional nasal swab 822; and [0768] test 302 (e.g., a diagnostic test), which is configured to test for the presence of a particulate in a specimen sample collected using the techniques described herein; test 302 is described hereinabove with reference to FIG. 12.

[0769] Nasal swab 822 comprises a swab tip 838 that comprises an absorbent material 844 for collecting the specimen sample. As used in the present application, including in the claims and Inventive Concepts, an “absorbent material” is a material that collects and retains a specimen sample, such as because of the physical or chemical structure and/or properties of the material (such as, for example, adhesive properties), and/or the structure formed by the material. For example, the fibers of synthetic flocking are arranged to be absorbent, even if the fibers in isolation from one another would not be absorbent. For example, swab tip 838 may be a flocked swab tip (e.g., comprising nylon), a rayon swab tip, a cotton swab tip, a polyurethane foam swab tip, a polyethylene terephthalate (PET) swab tip, a polyester swab tip, a foam swab tip (e.g., comprising polyurethane) (e.g., an open-cell foam swab), a sponge swab tip (e.g., comprising cellulose), a brush swab tip (e.g., similar to a cytology brush), and a nanofiber swab tip.

[0770] Reference is now made to FIG. 19, which is a schematic illustration of a testing kit 900, in accordance with an application of the present invention. Testing kit 900 comprises: [0771] a nasal dispenser (containing nasal wash fluid 26), such as conventional nasal dispenser 820, as shown, or nasal wash fluid dispenser 20, not shown in FIG. 19, but shown in FIG. 2B; [0772] nasal swab 822; and [0773] vial 402 and liquid 404 for bathing at least a portion of a specimen sample in the vial, the liquid selected from the group consisting of: a lysis buffer, saline solution (e.g., phosphate buffered saline (PBS)), and transport medium (e.g., universal transport medium or a viral transport medium).

[0774] Vial 402 and liquid 404 are described hereinabove with reference to FIG. 13.

[0775] Nasal swab 822 comprises swab tip 838 that comprises absorbent material 844 for collecting the specimen sample. For example, swab tip 838 may be a flocked swab tip (e.g., comprising nylon), a rayon swab tip, a cotton swab tip, a polyester swab tip, a polyurethane foam swab tip, a polyethylene terephthalate (PET) swab tip, a foam swab tip (e.g., comprising polyurethane) (e.g., an open-cell foam swab), a sponge swab tip (e.g., comprising cellulose), a brush swab tip (e.g., similar to a cytology brush), and a nanofiber swab tip.

[0776] Testing kit 900 may be used, for example, for collecting a specimen sample from the subject and sending at least a portion of the specimen sample to a remote laboratory in the liquid in the vial, such as for performing Polymerase Chain Reaction (PCR) testing for particulate in the specimen sample. The particulate may be a virus, a bacterium, any of the other particulates described hereinabove, or any of the other biological materials described hereinabove.

[0777] Reference is now made to FIGS. 20A-B and 21A-D, which are schematic illustrations of respective configurations of a nasal sampling device 1020 for collecting a specimen sample, in accordance with respective applications of the present invention. FIGS. 20A-B show a nasal sampling device 1020A and FIGS. 21A-B show a nasal sampling device 1020B. Nasal sampling device 1020A and nasal sampling device 1020B are configurations of nasal sampling device 1020 and are identical to each other except as described hereinbelow.

[0778] Nasal sampling device 1020 comprises a dispensing container 1072 and a nasal applicator 1038 (which is sized and shaped to be insertable into an anterior nares of a nostril). Dispensing container 1072 contains nasal wash fluid 26; optionally, nasal wash fluid 26 has any of the properties of nasal wash fluid 26 described hereinabove with reference to FIG. 2B.

[0779] Nasal applicator 1038 is shaped so as to define (i) one or more nozzles 1040, (ii) a proximal opening 1042, and (iii) one or more channels 1044 within nasal applicator 1038 that connect the one or more nozzles 1040 in fluid communication with proximal opening 1042. For example, nasal applicator 1038 may be shaped so as define exactly one dispensing opening 1040, which may, for example, be located at a distal end of nasal applicator 1038, such as shown. Alternatively, nasal applicator 1038 may be shaped so as define a plurality of nozzles 1040, which may include, for example, openings facing laterally (such as shown in FIG. 21D) and, optionally, an opening at the distal end of nasal applicator 1038.

[0780] At least a portion of an external surface of nasal applicator 1038 comprises an absorbent material 1044 for collecting the specimen sample. For example, absorbent material 1044 may comprise flocking (e.g., comprising nylon), rayon, cotton, polyester, polyurethane foam, polyethylene terephthalate (PET), foam (e.g., comprising polyurethane) (e.g., an open-cell foam), sponge (e.g., comprising cellulose), a brush (e.g., similar to a cytology brush), and nanofibers.

[0781] For some applications, nasal applicator 1038 is shaped similarly to a conventional nasal swab.

[0782] In the configurations shown in FIGS. 20A-B and 21A-B, an entirety of the external surface of nasal applicator 1038, other than the portion that defines the one or more nozzles 1040, comprises absorbent material 1044. In the configurations shown in FIGS. 21C-D, only a portion of the external surface of nasal applicator 1038 comprises absorbent material 1044.

[0783] For some applications, nasal applicator 1038 may implement any of the nozzles described hereinabove with reference to FIGS. 1, 2A-C, 3A-B, 6, 7A-K, 8A-C, 9A-F, 10, and/or 11, including the angles, offset angles, spray angles, number of nozzles, and orientations of the nozzles.

[0784] Nasal applicator 1038 is disposed distal to dispensing container 1072, with proximal opening 1042 of nasal applicator 1038 in fluid communication with dispensing container 1072. Nasal sampling device 1020 is configured to intranasally dispense nasal wash fluid 26 from dispensing container 1072 into nasal cavity 22 of the one or more nozzles 1040 of nasal applicator 1038.

[0785] For some applications, nasal sampling device 1020 comprises a pump 1046 that is configured to dispense nasal wash fluid 26.

[0786] For some applications, nasal sampling device 1020 is configured to dispense nasal wash fluid 26 by manual compression of dispensing container 1072.

[0787] For some applications, nasal sampling device 1020 comprises an atomizer that is configured to dispense nasal wash fluid 26.

[0788] For some other applications, nasal sampling device 1020 comprises a pressurized aerosol generator that is configured to dispense nasal wash fluid 26.

[0789] Reference is made to FIGS. 20A-B. For some applications, nasal sampling device 1020A further comprises a tube 1048, which physically connects a proximal end 1050 of nasal applicator 1038 to dispensing container 1072. Proximal opening 1042 of nasal applicator 1038 is in fluid communication with dispensing container 1072 via tube 1048.

[0790] Optionally, tube 1048 extends distally through at least a portion of nasal applicator 1038, so as to define at least a portion of the one or more channels 1044 within nasal applicator 1038, such as shown in FIGS. 20A-B.

[0791] For some applications, tube 1048 physically connects proximal end 1050 of nasal applicator 1038 to dispensing container 1072 such that a distance D between proximal end 1050 of nasal applicator 1038 and a distal-most point 1052 of dispensing container 1072 is at least 1 cm, no more than 13 cm, and/or between 1 and 13 cm.

[0792] Reference is made to FIGS. 21A-B. For some applications, proximal end 1050 of nasal applicator 1038 of nasal sampling device 1020B is in direct physical contact with dispensing container 1072.

[0793] For some applications, proximal end 1050 of nasal applicator 1038 is fixed in the direct physical contact with dispensing container 1072. For other applications, proximal end 1050 of nasal applicator 1038 is removably coupled in the direct physical contact with dispensing container 1072.

[0794] Reference is made to FIGS. 20A-B and 21A-B. For some applications, nasal applicator 1038 is removable from dispensing container 1072.

[0795] Reference is still made to FIGS. 20A-B and 21A-B. For some applications, nasal sampling device 1020 comprises a user control 1054 that is coupled to dispensing container 1072 and configured to cause dispensing container 1072 to dispense nasal wash fluid 26. For some of these applications, proximal end 1050 of nasal applicator 1038 is fixed to a distal portion 1056 of user control 1054, and at least distal portion 1056 of user control 1054 is removable from dispensing container 1072, so as to remove nasal applicator 1038 from dispensing container 1072. Either the entirety of user control 1054 is removable from dispensing container 1072, or only a portion of user control 1054 is removable from dispensing container 1072, while another portion of user control 1054 remains coupled to dispensing container 1072.

[0796] Reference is now made to FIG. 22, which is a schematic illustration of the use of nasal sampling device 1020 for collecting a specimen sample from nasal cavity 22, in accordance with an application of the present invention. Although by way of illustration FIG. 22 shows nasal sampling device 1020A, described hereinabove with reference to FIGS. 20A-B, nasal sampling device 1020B, described hereinabove with reference to FIGS. 21A-B, may be used in the same manner. As illustrated in FIG. 22, nasal sampling device 1020 includes exactly one nozzle 1040.

[0797] As shown in FIG. 22, nasal applicator 1038 of nasal sampling device 1020 is introduced into nostril 28, and thereafter nasal wash fluid 26 is intranasally dispensed, by the subject or by somebody else, into nasal cavity 22 from the one or more nozzles 1040, by introducing nasal wash fluid 26 into proximal opening 1042 (from dispensing container 1072). Typically, nasal applicator 1038 is introduced into nostril 28 no more deeply than an anterior naris 1060 of nostril 28.

[0798] Thereafter, the specimen sample is collected from nasal cavity 22 by contacting (typically, rubbing) a wall 1058 of anterior naris 1060 with absorbent material 1044 of nasal applicator 1038, by the subject or by somebody else. Typically, no portion of nostril 28 deeper than anterior naris 1060 is contacted by absorbent material 1044 of nasal applicator 1038. Typically contacting wall 1058 of anterior naris 1060 with absorbent material 1044 of nasal applicator 1038 comprises rubbing wall 1058 of anterior naris 1060 with absorbent material 1044 of nasal applicator 1038. Typically, nasal applicator 1038 is not removed from and reinserted into nostril 28 between intranasally dispensing nasal wash fluid 26 and collecting the specimen sample. Alternatively, nasal applicator 1038 is removed from and reinserted into nostril 28 between intranasally dispensing nasal wash fluid 26 and collecting the specimen sample.

[0799] The method may be performed in one or both of the nostrils. Optionally, the method is performed in combination with other sampling methods described herein, including, but not limited to gargling after dispensing nasal wash fluid 26.

[0800] For some applications, collecting the specimen sample from nasal cavity 22 comprises collecting the specimen sample without using suction. Alternatively, suction is used to collect the specimen sample.

[0801] For some applications in which nasal applicator 1038 is removable from dispensing container 1072, after the specimen sample is collected, nasal applicator 1038 is removed from dispensing container 1072. Optionally, nasal applicator 1038 is inserted into a liquid, such as a lysis buffer, saline solution (e.g., phosphate buffered saline (PBS)), transport medium (e.g., universal transport medium or a viral transport medium), or a composition comprising a detergent and a buffering agent for stabilizing RNA (e.g., distributed by DNA Genotek Inc., a subsidiary of OraSure Technologies, Inc. (Bethlehem, Pa., USA), for example, ORAcollect®⋅RNA Saliva Collection Device for SARS CoV-2). The liquid is optionally contained in collection receptacle 40, described hereinabove with reference to FIG. 2D.

[0802] For some applications, before the specimen sample is collected using the techniques described with reference to FIG. 22: [0803] before nasal wash fluid 26 is intranasally dispensed, the subject (typically while sitting or standing) tilts back his or her head (typically until the neck is fully extended) and holds the head tilted back while nasal wash fluid 26 is intranasally dispensed, or [0804] after nasal wash fluid 26 is intranasally dispensed, the subject tilts back his or her head farther back than while intranasally dispensing the nasal wash fluid and farther back than a neutral position.

[0805] In all of these cases, the tilting back may cause nasal wash fluid 26 to loosen biological material in a more posterior (deeper) portion of nasal cavity 22 and/or nasopharynx 32.

[0806] Optionally, after the subject tilts his or her head back and after nasal wash fluid 26 is intranasally dispensed, the subject tilts forward his or her head to a neutral or a tilted-forward position; this backward-followed-by-forward tilting may help move biological material anteriorly (i.e., toward the nostril) within nasal cavity 22.

[0807] Optionally, the method further comprises, while holding his or her head tilted back, waiting for a period of time between (a) concluding intranasally dispensing nasal wash fluid 26 into the nasal cavity and (b) tilting his or her head forward to the neutral or the tilted-forward position, the period of time at least 2 seconds (e.g., at least 5 seconds, such as at least 10 seconds, e.g., at least 15 seconds), no more than 60 seconds (e.g., no more than 30 seconds), and/or between 2 seconds (e.g., 5 seconds, such as at least 10 seconds, e.g., at least 15 seconds) and 60 seconds (e.g., 30 seconds).

[0808] For some applications, the subject tilts his or her head forward to the neutral or the tilted-forward position after the subject senses (e.g., by feeling and/or tasting) the nasal wash fluid in oropharynx 30. Optionally, nasal wash fluid 26 comprises a tastant, for example, menthol flavoring.

[0809] Alternatively, for some applications, nasal wash fluid 26 is intranasally dispensed into nasal cavity 22 (either continuously or in a plurality of pulses) until the subject senses (e.g., by feeling and/or tasting) nasal wash fluid 26 in oropharynx 30. Optionally, nasal wash fluid 26 comprises a tastant, for example, menthol flavoring.

[0810] Optionally, the method further comprises waiting for a period of time between (a) concluding intranasally dispensing nasal wash fluid 26 into the nasal cavity and (b) collecting the specimen sample, the period of time at least 2 seconds (e.g., at least 5 seconds, such as at least 10 seconds), no more than 30 minutes (e.g., no more than 15 minutes), and/or between 2 seconds (e.g., 5 seconds, such as at least 10 seconds) and 30 minutes (e.g., 15 minutes).

[0811] For some applications, nasal wash fluid 26 is intranasally dispensed as one or more types of fluid discharge selected from the group consisting of: a spray, a mist, a pressurized aerosol, steam, and an atomized fluid. For some applications, nasal wash fluid 26 is intranasally dispensed as a fluid stream.

[0812] For some applications, nasal wash fluid 26 is dispensed into each of one or both nostrils 28 in a total volume of between 0.15 and 1 mL, such as between 0.15 and 0.5 mL, or between 0.25 and 1 mL, such as between 0.25 and 0.5 mL. Optionally, nasal wash fluid 26 is intranasally dispensed in a plurality of pulses per nostril (e.g., between 3 and 5 pulses per nostril), each having a volume of between 0.05 and 0.2 mL, for example, between 0.05 and 0.15 mL, e.g., 0.1 mL. (If nasal wash fluid 26 is dispensed into both nostrils 28, the volumes above would be doubled.) Dispensing such relatively small volumes reduces the likelihood of nasal wash fluid 26 washing out the biological material of interest from the nostrils.

[0813] In some applications, the specimen samples collected using the collection techniques described herein are tested for the presence of a particulate using one or more of the techniques described in the patent applications incorporated hereinbelow by reference, or a technique known in the art. The particulate may be a virus (e.g., an Influenza virus or SARS-CoV-2), a bacterium (e.g., Streptococcus bacterium), any of the other particulates described hereinabove, or any of the other biological materials described hereinabove. For example, the particulate may be tested for the presence of a particulate using nucleic acid amplification, such as PCR, e.g., qPCR, and/or by performing an immunoassay, such as a lateral flow immunoassay, e.g., a chromatographic digital immunoassay, or by performing a rapid molecular test, for example one that uses a real-time loop mediated amplification reaction, such as the Lucira COVID-19 All-In-One Test Kit, or a NEAR technology, such as the ID NOW™, or a molecular test kit manufactured by Visby. Further alternatively, the particulate may be tested the presence of a particulate using a CRISPR-based diagnostic test, an ELISA diagnostic test, or a spectroscopy-based diagnostic test.

[0814] In an application of the present invention, a testing kit is provided that comprises nasal sampling device 1020 and test 302 (e.g., a diagnostic test), which is configured to test for the presence of a particulate in a specimen sample collected using the techniques described herein. Test 302 is described hereinabove with reference to FIG. 12.

[0815] In an application of the present invention, a testing kit is provided that comprises nasal sampling device 1020, vial 402, and liquid 404 for bathing at least a portion of a specimen sample in the vial, the liquid selected from the group consisting of: a lysis buffer, saline solution (e.g., phosphate buffered saline (PBS)), and transport medium (e.g., universal transport medium or a viral transport medium). Vial 402 and liquid 404 are described hereinabove with reference to FIG. 13. The testing kit may be used, for example, for collecting a specimen sample from the subject and sending at least a portion of the specimen sample to a remote laboratory in the liquid in the vial, such as for performing Polymerase Chain Reaction (PCR) testing for particulate in the specimen sample. The particulate may be a virus, a bacterium, any of the other particulates described hereinabove, or any of the other biological materials described hereinabove.

[0816] Reference is now made to FIGS. 23A and 23B, which are schematic illustrations of kits that comprise portions of nasal sampling devices 1020A and 1020B, respectively, and vial 402, in accordance with respective applications of the present invention. In these configurations, distal portion 1056 of user control 1054 is configured, after removal from dispensing container, such as described hereinabove with reference to FIGS. 20B and 21B, to be coupled to the opening of vial 402, such that nasal applicator 1038 is disposed within vial 402 and distal portion 1056 serves as a cap for vial 402. Vial 402 contains liquid 404, which bathes at least a portion of nasal applicator 1038 within vial 402. It is noted that distal portion 1056, serving as the cap, generally does not leak through the one or more channels 1044 and proximal opening 1042 of nasal applicator 1038, because proximal opening 1042 is too small to allow leakage.

[0817] Typically, after nasal applicator 1038 soaks in liquid 404, a portion of liquid 404 is removed for testing for a particulate, such as by removing the cap comprising distal portion 1056, and optionally coupling a conventional dropper cap to the vial and dispensing several drops of liquid 404, such as is known in the art.

[0818] In an application of the present invention, a testing kit is provided that comprises nasal sampling device 1020, vial 402, and liquid 404 for bathing at least a portion of a specimen sample in the vial, the liquid selected from the group consisting of: a lysis buffer, saline solution (e.g., phosphate buffered saline (PBS)), and transport medium (e.g., universal transport medium or a viral transport medium). test 302 (e.g., a diagnostic test), which is configured to test for the presence of a particulate in a specimen sample collected using the techniques described herein. Test 302 is described hereinabove with reference to FIG. 12, and vial 402 and liquid 404 are described hereinabove with reference to FIG. 13. The testing kit may be used, for example, for collecting a specimen sample from the subject and sending at least a portion of the specimen sample to a remote laboratory in the liquid in the vial, such as for performing a molecular test, such as Polymerase Chain Reaction (PCR) testing or isothermal testing for particulate in the specimen sample. The particulate may be a virus, a bacterium, any of the other particulates described hereinabove, or any of the other biological materials described hereinabove.

[0819] Reference is now made to FIGS. 24A and 24B, which are schematic illustrations of a testing kit 1100 for testing a specimen sample, in accordance with an application of the present invention. Testing kit 1100 comprises: [0820] a vial 1102; [0821] a liquid reagent 1104 (such as a lysing buffer, e.g., a lysing buffer configured to be used with an immunoassay); liquid reagent 1104 may be provided in vial 1102 or in a separate container; [0822] a cap 1108, which is configured to be sealingly coupled, at a distal side 1110 thereof, to a proximal opening 1112 defined by vial 1102; [0823] an elongate test-strip holder 1114, which is coupled to a proximal side 1116 of cap 1108; and [0824] a lateral flow test strip 1106, which is at least partially disposed within elongate test-strip holder 1114, at least partially visible from outside test-strip holder 1114 (for example, test-strip holder 1114 may comprise a transparent material and/or a window, which may optionally include an optical element, such as a magnifying element).

[0825] Cap 1108 is shaped so as to define an aperture 1118 that is in fluid communication with (a) distal side 1110 of cap 1108 and (b) lateral flow test strip 1106 within elongate test-strip holder 1114.

[0826] Cap 1108 comprises an absorbent material 1120 that is disposed on distal side 1110 of cap 1108 and is in fluid communication (a) with lateral flow test strip 1106 via aperture 1118, and (b) when cap 1108 is sealing coupled to proximal opening 1112 of vial 1102, with an interior 1122 of vial 1102. For example, absorbent material 1120 may comprise a sponge. Absorbent material 1120 is configured to regulate and control the volume of liquid reagent 1104 that flows from vial 1102 to lateral flow test strip 1106.

[0827] For some applications, lateral flow test strip 1106 comprises a lateral flow immunoassay test strip.

[0828] For some applications, vial 1102 has a volume of at least 3 cc, no more than 5 cc, and/or between 3 and 5 cc. For some applications, liquid reagent 1104 has a volume of at least 0.2 cc, no more than 1 cc, and/or between 0.2 and 1 cc.

[0829] During use, the specimen sample is placed in liquid reagent 1104, such as by inserting a swab containing the specimen sample into liquid reagent 1104 in vial 1102. Cap 1108 is sealingly coupled to proximal opening 1112 of vial 1102. Vial 1102 is typically turned over such that liquid reagent 1104 comes in contact with absorbent material 1120 of cap 1108. Absorbent material 1120 brings a small portion of liquid reagent 1104 into contact with lateral flow test strip 1106. The results may be directly observed, as is conventional in the art, and/or analyzed by a chromatographic digital immunoassay, such as the BD Veritor™ system.

Experimental Results

[0830] The inventors performed a number of experiments using some of the techniques described herein.

[0831] Although many of the experiments described herein include testing for the presence of the SARS-CoV-2 virus, the inventors believe that the testing techniques used in these experiments would provide similar results for other viruses sharing one or more characteristics with SARS-CoV-2, such as infection of the nasal cavity and viral tropism. Such other viruses include, for example, Influenza viruses, and rhinoviruses.

[0832] For example, there is shared tropism in Influenza and SARS-CoV-2 infection. The expression of receptors used in cellular entry is one of the principal limitations on viral tropism, with SARS-CoV-2 and Influenza requiring Angiotensin-converting enzyme-2 (ACE2) and Sialic acid (SA) expression, respectively, to be permissive for infection. The replication of SARS-CoV-2 in upper respiratory tract (URT) tissues is attributed to the expression of ACE2 in olfactory and respiratory epithelial cells. Similarly, the permissiveness of human nasal epithelial cells to Influenza infection in vitro is believed to be due to the cell-surface expression of SA in ciliated epithelial cells. Moreover, SARS-CoV-2 and Influenza have been reported to have similar tissue tropism in the lungs and bronchus. The inventors therefore believe that sampling methods described herein, such as saline nasal spraying and swabbing or saline nasal spraying and gargling, which produce samples with higher SARS-COV-2 viral loads and/or amounts of infected cell debris than traditional sampling methods, would similarly be a more effective sampling procedure for the diagnosis and/or detection of Influenza or any other URT infection which reproduce in nasal epithelial cells, such as human rhinoviruses.

Experiments 1, 2, and 3

[0833] Experiments 1, 2, and 3 demonstrate that at least a portion of intranasally-dispensed nasal wash fluid can be recovered from oral cavity 36 using methods described herein. It is noted that these experiments were performed with conventional, commercially-available nasal dispensers, as described in detail below, rather than with the novel two-nozzle nasal wash fluid dispenser 20 described hereinabove, or the novel three-nozzle nasal wash fluid dispenser 220 described hereinabove, although such use may improve the results of the methods.

Materials fir Experiments 1, 2, and 3

[0834] Standard nasal wash saline solution (hereinbelow, the “standard nasal wash saline solution”): FLO Sinus Care Starter Kit, ENT Technologies Pty Ltd., Hawthorn East Victoria 3123, Australia. (The FLO Sinus Care Starter Kit is marketed and described as a nasal wash saline solution for sinus care.) [0835] Colored nasal wash fluid (hereinbelow, the “colored nasal wash fluid”): A proprietary blue colored nasal wash fluid was made by combining the standard nasal wash saline solution with a standard blue food coloring (made in the USA). [0836] Oral wash fluid (hereinbelow, the “oral wash fluid”): FLO Sinus Care Starter Kit, ENT Technologies Pty Ltd., Hawthorn East Victoria 3123, Australia. (The FLO Sinus Care Starter Kit is marketed and described as a nasal wash saline solution tier sinus care; the inventors of the present application have used it as the novel oral wash fluid described herein.) [0837] Standard nasal spray dispenser (hereinbelow, the “standard nasal spray dispenser”): Afrin Nasal Spray, distributed by MSD Consumer Care Inc., POB 377 Memphis, Tenn., USA, a subsidiary of Merck and co. Inc., White House Station, N.J., USA. The dispenser was emptied of all contents and washed out prior to use.

Methods for Experiments 1, 2, and 3

[0838] Setting up the standard nasal wash fluid syringe dispenser: Nasal wash fluid was transferred into a 5 mL syringe with a distally attached rubber tubing. The rubber tubing was introduced about 1 cm into nasal cavity 22 via one nostril 28 and was positioned along the floor of nasal cavity 22 so as to direct the nasal wash fluid a posterior direction 48 towards nasopharynx 32.

Experiment 1: Colored Nasal Wash Fluid

[0839] In Experiment 1, the nasal wash fluid syringe dispenser was set up with 3.0 mL of the prepared colored nasal wash fluid. [0840] The colored nasal wash fluid was dispensed as a fluid stream into nasal cavity 22 in a posterior direction 48 towards nasopharynx 32 by manually pressing the plunger of the syringe. [0841] Within a few seconds, the colored nasal wash fluid was felt in the throat, which is hypothesized to be the point in time at which the colored nasal wash fluid began draining from nasopharynx 32 into oropharynx 30. [0842] A specimen sample was then expressed out of oral cavity 36 by spitting out the contents of oral cavity 36 through anterior opening 34 of oral cavity 36.

Results of Experiment 1

[0843] The orally expressed specimen sample had a volume of about 0.5 to 0.6 mL and was confirmed to contain at least a portion of the intranasally dispensed colored nasal wash fluid by visually ascertaining a distinct blue color in the specimen sample.

Conclusions Regarding Experiment 1

[0844] At least a portion of an intranasally dispensed colored nasal wash fluid was recovered in orally expressed specimen sample.

Experiment 2: Testing Different Volumes

[0845] In Experiment 2, different volumes of nasal wash fluid were tested. [0846] For each experiment, the nasal wash fluid syringe dispenser was set up with different volumes ranging from 0.5 to 5.0 mL of nasal wash fluid consisting of the standard nasal wash saline solution. Each volume was tested once or multiple times. [0847] For each experiment, a volume of nasal wash fluid was dispensed as a fluid stream into nasal cavity 22 in a posterior direction 48 towards nasopharynx 32 by manually pressing the plunger of the syringe, while head was tilted back about 15-30 degrees. [0848] Within a few seconds, the nasal wash fluid was felt and/or tasted in the throat, which is hypothesized to be the point in time at which the nasal wash fluid began draining from nasopharynx 32 into oropharynx 30. [0849] When possible (as explained below), after draining into oropharynx 30, the nasal wash fluid was gargled. [0850] When possible (as explained below), a specimen sample was then expressed out of oral cavity 36 by spitting out the contents of oral cavity 36 through anterior opening 34 of oral cavity 36.

Results of Experiment 2

[0851] When 0.5 mL was intranasally dispensed (tested 4 times): Fluid was felt in the throat but not enough to be expressed by spitting out [0852] When 1.0 mL was intranasally dispensed (tested 7 times): in some cases, fluid was felt in the throat, but there was not enough to be expressed by spitting out. In other cases, between 0.13 and 0.22 mL of specimen sample was expressed by spitting out. For all cases of 1.0 mL dispensed, it was not possible to gargle. [0853] When 2.0 mL was intranasally dispensed (tested 2 times): It was possible to perform a weak gargle, and between 0.3 and 0.4 mL of specimen sample was expressed by spitting out. [0854] When 3.0 mL was intranasally dispensed (tested 4 times): It was possible to perform a weak gargle, and between 0.55 and 1.2 mL of specimen sample was expressed by spitting out. [0855] When 4.0 mL was intranasally dispensed (tested 2 times): it was possible to perform a gargle, and about 1.0 mL of specimen sample was expressed by spitting out. [0856] When 5.0 mL was intranasally dispensed (tested 2 times): It was possible to perform a gargle, and between 1.2 and 2.1 ml of specimen sample was expressed by spitting out.

Conclusions Regarding Experiment 2

[0857] Specimen samples can be orally expressed when more than about 1 mL of nasal wash fluid is intranasally dispensed. Nasal wash fluid can be gargled prior to orally expressing specimen sample when more than about 2 ml of nasal wash fluid is intranasally dispensed. However, the inventors hypothesize that dispensing a smaller volume of as low as 0.2 mL into nasal cavity 22 may be adequate for draining to oropharynx 30 if the fluid is dispensed at a higher pressure and/or better aimed toward nasopharynx 32 than in the experiments performed.

Experiment 3: Nasal Wash Fluid+Oral Wash Fluid Gargled

[0858] The standard nasal spray dispenser was used to dispense a single pulse of nasal wash fluid consisting of the standard nasal wash saline solution into one nostril 28 in the posterior direction 48 within nasal cavity 22. [0859] Then, after the nasal wash fluid was felt and/or tasted in the throat, 5 mL of the oral wash fluid was added to oral cavity 36 vial anterior opening 34 of oral cavity 36. [0860] The nasal wash fluid and oral wash fluid were gargled together and then expressed out of anterior opening 34 of oral cavity 36 as a first specimen sample by spitting out. [0861] After spitting out the first specimen sample, an additional 5 ml, of oral wash fluid was added to oral cavity 36 vial anterior opening 34 of oral cavity 36 without dispensing any additional nasal wash fluid. [0862] The oral wash fluid was gargled and then expressed out of anterior opening 34 of oral cavity 36 as a second specimen sample by spitting out. [0863] Volumes of both specimen samples were measured.

Results of Experiment 3

[0864] Volume of first specimen sample (gargled nasal wash fluid+oral wash fluid): 4.56 mL [0865] Volume of second specimen sample (gargled oral wash fluid): 4.3 ml. [0866] Volume of nasal wash fluid dispensed in a single pulse: 1.98 mL
Conclusions regarding Experiment 3

[0867] There was a slight increase in specimen sample volume when the nasal wash fluid is dispensed compared to gargling with oral fluid alone, indicating that at least a portion of the nasal wash fluid was washed into oropharynx 30 and was expressed out of oral cavity 36.

Experiments 4 and 5

[0868] Experiments 4 and 5 demonstrate that at least a portion of biological material derived from nasal cavity 22 and/or nasopharynx 32 can be washed into oropharynx 30 by the intranasally-dispensed nasal wash fluid and can then be recovered from oral cavity 36 using methods described herein. It is noted that these experiments were performed with conventional, commercially-available nasal dispensers, as described in detail below, rather than with the novel two-nozzle nasal wash fluid dispenser 20 described hereinabove, or the novel three-nozzle nasal wash fluid dispenser 220 described hereinabove, although such use may improve the results of the methods.

Materials fir Experiments 4 and 5

[0869] Standard nasal wash saline solution (hereinbelow, the “standard nasal wash saline solution”): PLO Sinus Care Starter Kit, ENT Technologies Pty Ltd., Hawthorn East Victoria 3123, Australia. [0870] Standard metered nasal mist dispenser (hereinbelow, the “standard metered nasal mist dispenser”): NasaCort, distributed by Chattem Sanofi Company, Chattanooga, Tenn., USA. The dispenser was emptied of all contents and washed out prior to use.

Experiment 4: Asymptomatic Subject—Microscopic Analysis of Specimen Sample

[0871] The expressed specimen sample collected from Experiment 1, described hereinabove, was centrifuged at 3500 RPM for 10 minutes. [0872] Supernatant was discarded and pellet was isolated. A portion of the pellet was mounted on a glass slide and stained using Gram staining to enable viewing of nuclei and cellular shape. [0873] The slides were analyzed by light microscopy.

Results of Experiment 4

[0874] The slides showed large quantity of squamous epithelial cells, which are morphologically square in shape and are known in the art to be present in the oral cavity. The slides also showed minimal quantity of columnar epithelial cells, which are morphologically elongated and rectangular in shape and are known in the art to be present in the nasal cavity and not present in the oral cavity.

Conclusions Regarding Experiment 4

[0875] Microscopic analysis of orally expressed specimen sample shows cells of nasal origin, confirming that biological material was washed from nasal cavity 22 into oropharynx 30 to be expressed out of oral cavity 36 by spitting out.

Experiment 5: Symptomatic Subject—Microscopic Analysis of Specimen Sample

[0876] Experiment 5 was performed on a subject suffering from seasonal allergic rhinitis. [0877] The standard metered nasal mist dispenser was used to dispense nasal wash fluid consisting of the standard nasal wash saline solution intranasally according to the following procedure: [0878] (1) Spray twice in the first nostril: once superoposteriorly and once posteriorly, [0879] (2) Spray twice in the second nostril, once superoposteriorly and once posteriorly. [0880] (3) Repeat steps (1) and (2) for a total of 8 sprays. [0881] Within a few seconds, the nasal wash fluid was felt and/or tasted in the throat, which is hypothesized to be the point in time at which the nasal wash fluid began draining from nasopharynx 32 into oropharynx 30. [0882] A specimen sample was then expressed out of oral cavity 36 by spitting out the contents of oral cavity 36 through anterior opening 34 of oral cavity 36. The volume of the specimen sample that was collected was about 2 mL. [0883] The expressed specimen sample was centrifuged at 3000 RPM for 10 minutes. [0884] Supernatant was discarded and pellet was isolated, A portion of the pellet was mounted on a glass slide and stained using Gram staining to enable viewing of nuclei and cellular shape. [0885] The slides were by light microscopy.

Results of Experiment 5

[0886] The slides showed large quantity of columnar epithelial cells, which are morphologically elongated and rectangular in shape and are known in the art to be present in the nasal cavity and not present in the oral cavity. The slides also showed minimal quantity of squamous epithelial cells, which are morphologically square in shape and are known in the art to be present in the oral cavity. The slides also showed eosinophils, which are morphologically distinct due to the presence of distinct large granules and are known in the art to be present in the nasal cavity and not present in the oral cavity. Eosinophils were present both as intact cells and as degranulated cells with free granules.

Conclusions Regarding Experiment 5

[0887] Microscopic analysis of orally expressed specimen sample shows high quantity of cells of nasal origin, confirming that biological material was washed from nasal cavity 22 into oropharynx 30 to be expressed out of oral cavity 36 by spitting out.

[0888] Asymptomatic subjects with minimal nasal secretions appear to have fewer epithelial cells recovered from the nasal cavity than symptomatic subjects with nasal secretions. The inventors hypothesize that this may be due to symptomatic subjects having comparatively higher quantity of desquamated cells and/or damaged cells that are more easily sloughed off and/or dislodged by the nasal wash fluid.

Experiment 6

[0889] Experiment 6 demonstrates that at least a portion of intranasally-dispensed nasal wash fluid can be recovered from oral cavity 36 using the novel methods and the two-nozzle nasal wash fluid dispenser 20 described herein. It is noted that this experiment was performed with a novel two-nozzle nasal wash dispenser similar to the two-nozzle nasal wash fluid dispenser 20 described herein, rather than with the novel three-nozzle nasal wash fluid dispenser 220 described hereinabove, although such use may improve the results of the methods.

Materials for Experiment 6

[0890] Standard nasal wash saline solution (hereinbelow, the “standard nasal wash saline solution”): FLO Sinus Care Starter Kit, ENT Technologies Pty Ltd., Hawthorn East Victoria 3123, Australia. [0891] Colored nasal wash fluid (hereinbelow, the “colored nasal wash fluid”): A proprietary blue colored nasal wash fluid was made by combining the standard nasal wash saline solution with a standard blue food coloring (made in the USA). [0892] Oral wash fluid (hereinbelow, the “oral wash fluid”): FLO Sinus Care Starter Kit, ENT Technologies Pty Ltd., Hawthorn East Victoria 3123, Australia. (The FLO Sinus Care Starter Kit is marketed and described as a nasal wash saline solution for sinus care; the inventors of the present application have used it as the novel oral wash saline solution described herein.) [0893] Two-nozzle nasal spray dispenser (hereinbelow, the “two-nozzle nasal spray, dispenser”): a custom-manufactured nasal spray dispenser comprising a tubular applicator similar to tubular applicator 64, described hereinabove, that was attached to the distal end of a standard syringe for measured fluid dispensing. The tubular applicator was shaped so as to define two nozzles that enable fluid to be dispensed in two directions simultaneously. The first nozzle directed fluid in the posterior direction within the nasal cavity, and the second nozzle directed fluid in the superoposterior direction within the nasal cavity.

Methods for Experiment 6

[0894] Setting up the two-nozzle nasal spray dispenser: 0.4 mL of the colored nasal wash fluid was transferred into a 2.5 ml, syringe with the two-nozzle nasal spray tip attached to the standard tapered exit port of the syringe.
Experiment 6: colored Nasal Wash Fluid [0895] In Experiment 6, each nostril was sprayed with 0.4 mL of the colored nasal wash fluid using the two-nozzle nasal spray dispenser, and the subject sniffed the nasal wash fluid further into the nasal cavity after the nasal wash fluid was dispensed into the nostril. [0896] After a few seconds, once at least a portion of the nasal wash fluid was felt draining into oropharynx 30, 4 mL of the colorless oral wash fluid was dispensed into oral cavity 36 through anterior opening 34 of oral cavity 36, and gargled for approximately 10 seconds. [0897] A specimen sample was then expressed out of oral cavity 36 into CUP A by spitting out the contents of oral cavity 36 through anterior opening 34 of oral cavity 36. [0898] Each nostril was then sprayed with an additional 0.4 mL of colorless standard nasal wash saline solution using the two-nozzle nasal spray dispenser. [0899] After a few seconds, once at least a portion of the second nasal wash fluid was felt draining into oropharynx 30, and additional 4 mL of the colorless oral wash fluid was dispensed into oral cavity 36 through anterior opening 34 of oral cavity 36, and gargled for approximately 10 seconds. [0900] A second specimen sample was then expressed out of oral cavity 36 into CUP B by spitting out the contents of oral cavity 36 through anterior opening 34 of oral cavity 36,

Results of Experiment 6

[0901] CUP A: Observed blue color in sample. Sample volume was 3.68 mL. [0902] CUP B: Observed blue color in sample. Sample volume was 4.88 mL.

Conclusions Regarding Experiment 6

[0903] At least a portion of an intranasally dispensed colored nasal wash fluid was recovered in orally expressed specimen sample when using a small volume of nasal wash fluid. A two-nozzle nasal spray is functional and may increase the sample yield. Residual color from the first nasal spray was recoverable when procedure was repeated, which indicates the possibility that repeating the procedure may increase the sample yield.

Experiment 7

[0904] Experiment 7 provides evidence that viral particles can be captured and detected on filters having pore sizes substantially larger than the viral diameter when filtering samples such as gargled fluid, nasal washes, and saliva. The inventors hypothesize that filters retain mucus and other large or aggregated salivary components, which in turn contain viral particles, and/or that the viral particles adhere e.g., directly adhere) to the material of the large-pore filters. It is noted that this experiment was performed with conventional, commercially-available nasal dispensers, as described in detail below, rather than with the novel two-nozzle nasal wash fluid dispenser 20 described hereinabove, or the novel three-nozzle nasal wash fluid dispenser 220 described hereinabove, although such use may improve the results of the methods.

Materials for Experiment 7

[0905] Standard saline solution for nasal and oral wash (hereinbelow, the “standard nasal and oral wash saline solution): 0.9% w/v Sodium Chloride. Catalog Number 364-2828. B. Braun Melsungen AG. Melsungen, Germany. [0906] Nasal spray bottle (hereinbelow, the “standard nasal spray bottle”): Nasal Sprayer 0.1 mL dosage. Catalog number 0591-0410-18. Gil Pack Pharmaceutical Packaging. Ashdod, Israel. [0907] PCR Testing: Sent to external laboratory. Roche Cobus 4800. Two unspecified targets used. [0908] Immunoassay Kits (hereinbelow, the “rapid immunoassay”): One Step Test Device. SARS-CoV-2 Antigen Rapid Test. Nantong Egens Biotechnology Co., Ltd. China. [0909] Transport Medium (hereinbelow, “UTM”): Sample Collection Vial. Universal Transport Medium (UTM) 3 mL. Reference Number BM-801. NovaMed Ltd. Jerusalem, Israel. [0910] Polyester Filters: Polyester Non-Woven Colorful Felt. Ningbo Home-Dollar Imp. & Exp. Company. China. Cut into 25 mm diameter circles. For pore size, see below immediately following Table 6. [0911] Grade C Filters: Glass Fiber Filters, Grade C, 25 mm. 1.2 micron nominal pore size. C2500. Sterlitech. Kent, Wash. USA. [0912] Grade F Filters: Glass Fiber Filters, Grade F, 25 mm. 0.7 micron nominal pore size. F2500. Sterlitech. Kent, Wash. USA. [0913] PES Filters: Polyethersulfone (PES) Membrane Filters, 0.03 micron, 25 mm. PES00325100. Sterlitech. Kent, Wash. USA.

Methods for Experiment 7

[0914] One subject confirmed positive for SARS-CoV-2 infection was tested over the course of a 4-day period. [0915] Combined Nasal Spray & Gargle Procedure (hereinbelow, the “N&G Procedure”): The standard nasal and oral wash saline solution was sprayed 3-5 times (0.1 mL per spray, for a total of 0.3-0.5 mL) into each nostril with the standard nasal spray bottle until fluid was felt by the subject in the back of the throat. The subject sniffed deeply after spraying was complete. The subject then introduced about 5 mL of the standard nasal and oral wash saline solution via anterior opening 34 of oral cavity 36 and gargled the saline solution for about 15 seconds. The subject then spit out the liquid (hereinbelow, the “N&G liquid specimen sample”). [0916] Gargle Alone Procedure (hereinbelow, the “G Alone Procedure”): The subject introduced about 5 mL of the standard nasal and oral wash saline solution via anterior opening 34 of oral cavity 36 and gargled the saline solution for about 15 seconds. The subject then spit out the liquid (hereinbelow, the “G Alone liquid specimen sample”). The nasal and oral wash saline solution was not introduced into the nasal cavity in this procedure. [0917] Filtration procedure: Prior to filtration, the N&G and G Alone liquid specimen samples were swabbed and either tested by rapid immunoassay or placed into UTM for PCR testing. The N&G and G Alone liquid specimen samples were then filtered through a syringe containing one or two polyester filters, one or two grade C filters, and one grade F filter, separated by spacers. The intermediary filtrates were then filtered through a second syringe containing one PES filter. The final filtrates were swabbed and either tested by rapid immunoassay or placed into UTM for PCR testing. All filters were removed and either tested by rapid immunoassay or placed into UTM for PCR testing. All filtrations were conducted using an air pressure pump at 1.4 ATM.

Results and Discussion of Experiment 7

[0918] Table 4 shows qualitative immunoassay results. The samples were taken on two consecutive days (“Day 1” and “Day 2”). All fluid samples were positive for SARS-CoV-2 prior to filtration and all final filtrates were negative. All filters tested positive for SARS-CoV-2. Taken together, the data indicate that filters cohesively retained the virus during filtration, indicating overall successful filtration.

[0919] As also can be seen in the comparison between the N&G and G Alone liquid specimen samples in Table 4, the N&G liquid specimens produced substantially more intense test lines on the immunoassay test strips than the G Alone liquid specimens for the fluid before filtration and all of the tested filters except the Grade C filter (which had the same intensity).

TABLE-US-00004 TABLE 4 Qualitative Immunoassay Results (Intensity of Test Lines on Test Strips on an Arbitrary Scale from 0 to 5) Day Day 1 Day 2 Nasopharyngeal and Throat Swab (Control) 5 5 Fluid Sample Type N&G N&G G Alone Swab of Fluid Before Filtration 0.5 4 2 Polyester Filter 3 1.5 1 Grade C Filter 2.5 1.5 1.5 Grade F Filter 1.5 3 1 PES Filter 1.5 1 0.5 Swab of Filtrate 0 0 0

[0920] Table 5 shows results of PCR tests of a single N&G liquid specimen sample based on two targets. The results are expressed in cycle threshold (Ct) values, as is standard in quantitate PCR. As is known in the quantitative PCR art, the Ct value is defined as the number of cycles of amplification (using PCR) required for the fluorescence of a PCR product (i.e., the target/amplicon) to be detected crossing a threshold that is above the background signal. Relatively lower Ct values indicate higher initial viral levels. The single N&G liquid specimen sample was taken on “Day 4,” i.e., two days after “Day 2” mentioned above.

[0921] As can be seen in Table 5, the fluid sample before filtration had significantly lower Ct values compared to the filtrate, indicating successful filtration, consistent with the immunoassay data presented above in Table 4.

TABLE-US-00005 TABLE 5 Qualitative PCR Results (Ct values) Target Target 1 Target 2 Day Day 4 Day 4 Nasopharyngeal Swab (Control) 27.71 28.12 Mid-turbinate Swab (Control) 29.81 30.24 Outer Nares Swab (Control) 31.25 32.02 Swab of Fluid Before Filtration 30.14 30.53 Polyester Filter 29.09 30.12 Grade C Filter 31.21 31.86 Grade F Filter 31.59 32.08 PES Filter 30.41 30.52 Swab of Filtrate 36.86 39.53

[0922] Table 6 shows the pore size of each filter. Only the PES filter has a pore size that is less than the SARS-CoV-2 diameter (which is generally believed to range between 0.05 and 0.2 microns). However, the PES filter did not show improved viral retention compared to other filters (Table 5). In fact, filter Ct values were not significantly different from each other, indicating that all of the filters retained the virus with similar efficiency (p=0.55, Mann-Whitney test).

TABLE-US-00006 TABLE 6 Pore Sizes of Filters Pore Size Greater or Less than Filter Type (microns) SARS-CoV-2 diameter Polyester 122.5 > Grade C 1.2 > Grade F 0.7 > PES 0.03 <

[0923] The 122.5 micron nominal pore size of the polyester filter was characterized using the bubble point test (i.e., largest pore diameter) using a capillary flow porometer, Model Number CFP-1500A with options E, X, M, and L, made by Porous Materials Inc. (PMI) and Galwick (PMI) as the wetting liquid, which has a fluid surface tension of 15.9 dynes/cm. (The bubble point pressure was 0.054 PSI.)

[0924] The polyester filter was also characterized as having a smallest pore diameter of 64.4 microns as determined by capillary flow capillary flow porometry, and a mean pore diameter of 75.5 microns as determined by capillary flow porometry, with a standard deviation of 66.6. The polyester filter was further characterized as having a distribution of pore size values skewed towards the smallest size, with over 40% of pores between 65 and 70 microns.

Conclusions Regarding Experiment 7

[0925] Filters having pores larger than the viral diameter nonetheless retain virus from gargled fluid, saliva, and other types of oral and/or nasal wash fluid. The large-pore filter likely concentrates the virus indirectly via retained or trapped biological materials such as saliva and/or mucus. This concentrated sample may improve test sensitivity compared to unfiltered samples and the large pore size enables high flow rate and inhibits filter clogging despite high sample viscosity.

Experiment 8

[0926] Some of the techniques described hereinabove comprise intranasally dispensing nasal wash fluid into the nasal cavity; thereafter, gargling, by the subject, the nasal wash fluid washed into the oropharynx; and thereafter, collecting a specimen sample that passed out of the anterior opening of the oral cavity. Experiment 8 provides evidence of the efficacy of these techniques for collecting specimen samples for detection of SARS-CoV-2 using quantitative PCR (qPCR).

[0927] In summary, Experiment 8 found that: [0928] the combination of nasal wash dispensing and gargling was 40% more likely to be positively identified using qPCR as containing SARS-CoV-2 compared to gargle-only samples, and 22% more likely to be positively identified using qPCR as containing SARS-CoV-2 compared to anterior nares swabs; [0929] the combination of nasal wash dispensing and gargling improved SARS-CoV-2 detection using qPCR by between 543% and 837% depending on the target gene compared to paired gargle samples; and [0930] concentration of nasal gargle samples by filtration further improved SARS-CoV-2 detection, using qPCR, by an addition 373% to 450% compared to unconcentrated nasal gargle samples.

Detailed Report of Experiment 8: A Novel and Non-Invasive Nasopharyngeal Self-Sampling Methodology for the Detection of SARS-CoV-2

Section 1: Aim of Study

[0931] 1. To define the effects of administering a mild nasal spray prior to sample procurement via gargling (hereinbelow, “Nasal Gargle”) on SARS-CoV-2 detection sensitivity. [0932] 2. To examine the efficacy of utilizing a hand-held concentration device in concentrating SARS-CoV-2 RNA in gargle samples.

Section 2: Pivotal Results

[0933] 1. Nasal Gargle was 40% more likely to be positively identified using qPCR as containing SARS-CoV-2 compared to gargle samples, and 22% more likely to be positively identified using qPCR as containing SARS-CoV-2 compared to anterior nares swabs. [0934] 2. Nasal Gargle improved SARS-CoV-2 detection using qPCR by between 543% and 837% depending on the target gene compared to paired gargle samples. [0935] 3. Concentration of Nasal Gargle samples by filtration further improved SARS-CoV-2 detection, using qPCR, by an additional 373% to 450% compared to unconcentrated nasal gargle samples.

Section 3: Background

[0936] SARS-CoV-2 diagnostics can be greatly expanded by a sampling technique that is both highly sensitive and allows self-collection. Multiple independent studies performed in Germany, India, British Columbia, and most recently, at the University of Arizona, have demonstrated that oropharyngeal gargle samples obtained by gargling 5 mL to 10 mL of saline are equivalent.sup.1,2,3, if not significantly more sensitive (>20%).sup.4 than nasopharyngeal (hereinbelow, “NP”) swabbing for SARS-CoV-2 detection. Gargle samples were also found to be more sensitive than other self-sampling techniques such as saliva collection when tested in parallel.sup.1. Furthermore, the FDA has recently approved the OraRisk COVID-19 RT-PCR test for the collection and processing of gargle samples under EUA for SARS-CoV-2 diagnosis.sup.5. However, despite NP swabbing and equivalent sampling techniques being the current gold-standard for SARS-CoV-2 detection, evidence suggests that these methods are highly variable and cause a concerning number of false-negative results in clinical settings.sup.6,7,8,9. Accordingly, we sought to improve the gargle sampling methodology, and to confirm a significant incremental benefit by coupling gargling with a swab-free non-invasive method for sampling the nasopharyngeal region, along with an additional step to further increase sensitivity by means of a novel concentration technique. We have successfully validated the underlying premise of these novel techniques.

Section 4: Rationale

[0937] Observed discrepancies between paired sampling procedures targeting variable regions in the upper respiratory tract suggest that the physiological localization and distribution of SARS-CoV-2 is heterogeneous in nature, and likely a contributing factor for noted variations in testing sensitivity between sampling methodologies.sup.9. Similarly, self-sampling techniques such as anterior nares swabbing have routinely underperformed when testing asymptomatic individuals.sup.10, indicating differential regional availability of SARS-CoV-2 in the upper respiratory tract between cohorts. Furthermore, saliva and anterior nares swabs have been found to be poor sample candidates for immunoassays, with heavily invasive NP swabbing performed by a trained medical professional preferred.sup.11, limiting the potentiality of an effective at-home diagnostic device. We have invented a robust and non-invasive sampling technique which simultaneously targets both the nasopharyngeal and oropharyngeal regions, in which nasal spray is administered for the sampling of virus-rich nasopharyngeal regions, which by post-nasal drip is accessible through an oropharyngeal gargle. We posited that by utilizing the combined Nasal Gargle as a sample type, patients that have viral replication primarily localized to specific sites which would be otherwise inaccessible to current self-sampling techniques such as saliva or gargle collection, or anterior nares swabbing alone, would be more readily detected, increasing the sensitivity of any downstream molecular or immunological tests performed. Moreover, we examined the additive effects of subsequent concentration of Nasal Gargle samples by filtration to further optimize SARS-CoV-2 detection.

Section 5: Methods

5.1 Sample Collection

[0938] In preparing gargle specimens, inpatients (n=21) in the coronavirus ward at Laniado Hospital (Netanya, Israel) were asked to gargle 5-10 mL of normal saline for approximately 9-15 seconds, and then spit the gargle fluid into a sterile collection receptacle. For the generation of Nasal Gargle specimens, the patients first sprayed 0.1 mL of normal saline 3-5 times into each nostril. Once the saline was felt in the back of the throat, patients were requested to sniff, followed by performing a gargle by introducing 5-10 mL of normal saline into the oral cavity via the anterior opening of the oral cavity and gargling for approximately 9-15 seconds, and then spitting the gargled fluid into a sterile collection receptacle. A flocked swab (3BY Ltd., Industrial Park Tefen, Israel, Ref: 3by-001) was waved through the Nasal Gargle or Gargle specimens for approximately 10 seconds, in a manner such that all internal surfaces of the collection receptacle were contacted by the swab. The swab was then placed into tubes containing 3 mL of Universal Transport Medium (UTM) (Biological Industries, Israel). Bilateral anterior nares swabbing (3BY Ltd., Industrial Park Tefen, Israel, Ref: 3by-001) was performed by hospital staff in parallel to gargle and Nasal Gargle sample collection (n=12). Anterior nares swabs were similarly placed into tubes containing 3 mL of UTM (Biological Industries, Israel). Tubes were left at room-temperature for up to 6 hours or stored at 4° C. for up to 72 hours prior to qPCR testing. For 3 of the 12 patients, Nasal Gargles were produced twice in succession. Pairwise comparisons between Nasal Gargle and swabbing or Nasal Gargle and gargle were performed with the first Nasal Gargle sample produced. Prior to concentration (described in Section 5.2) the paired Nasal Gargles were pooled together and then re-aliquoted. In addition to the anterior nares swab, for 5 of the 12 patients, mid-turbinate swabbing was performed by hospital staff prior to gargle sample production. The clinical study was approved by the Laniado Hospital investigation review board (IRB) Helsinki committee.

5.2 Concentration

[0939] 5-10 mL of collected Nasal Gargle was transferred into a hand-held concentration device using a serological pipette and passed through two of the Polyester Filters described and characterized hereinabove in the description of Experiment 7, while the two filters were disposed within the device. The filters were removed from the device, placed into a tube containing 3 mL of UTM (Biological Industries, Israel), and tubes were vortexed for 30-60 seconds. Tubes were left at room-temperature for up to 6 hours, or stored at 4° C. for up to 72 hours, prior to qPCR testing.

5.3 Molecular Testing

[0940] Prior to RNA extraction, tubes were vortexed for 30-60 seconds. RNA extraction was performed using a MagCore Viral nucleic acid extraction kit (High Sensitivity) (Ref: MVN400-06) on an Automated Nucleic Acid Extractor-MagCore Super (RBC Bioscience, Taiwan). The Allplex SARS-CoV-2 Assay (Seegene, Seoul, South Korea) was used for RNA detection on a CFX96 Real-Time System (BIO RAD, USA). Positive, presumptive positive, and negative SARS-CoV-2 scoring was preformed automatically by the assay's data analyzer, and both positive and presumptive positives were assigned as true positives. RNA extraction and qPCR were performed by the site study laboratory. The Wilcoxon Signed-Rank Test was used to determine statistical significance (p<0.05).

Section 6: Results

6.1: A Comparative Evaluation Between Nasal Gargle and Gargle Sampling Techniques

[0941] Paired Nasal Gargle and gargle samples were evaluated for the presence of SARS-CoV-2 by qPCR. Out of the 21 patients tested, 14 were found to be positive, with detection from Nasal Gargle outperforming that of gargle in 9 (RdRP/S) or 10 (E and N genes) of the 14 patients (FIG. 25A) based on relative ΔCt values. (As is known in the quantitative PCR art, the Ct value is defined as the number of cycles of amplification (using PCR) required for the fluorescence of a PCR product (i.e., the target/amplicon) to be detected crossing a threshold that is above the background signal. Relatively lower Ct values indicate higher initial viral levels.) Despite the small sample size, there was a statistically significant difference between the Nasal Gargle ΔCt values and the gargle ΔCt values for the E gene (p=0.02661) and the N gene (p=0.02148), with the RdRP/S gene approaching statistical significance (p=0.05225). Averaged differences between the two sample types showed a clear preference for Nasal Gargle, with ΔΔCt values of −2.442 (RdRP/S), −3.064 (E), or −3.006 (N) (FIG. 25B). This equates to a 5.434 to 8.365-fold (2.sup.−ΔΔCt) increase of viral material sampled by adding nasal spray prior to gargling. Moreover, the sensitivity of testing Nasal Gargle samples was 100%, compared to 71.4% when testing gargle samples (FIG. 25C).

[0942] Description of FIG. 25 (A Direct Comparison Between Nasal Gargle and Gargle): FIG. 25A) Paired Nasal Gargle (NG=left endpoint) and gargle (G=right endpoint) ΔCt values of target genes displayed after being normalized to a housekeeping gene tested in parallel. ΔCt values of NG tend to be less than relative G samples. Statistically significant differences in ΔCt values (p<0.05) between NG and G are indicated with an asterisk (*). FIG. 25B) The average difference between NG and G ΔCt values range from −2.48 to −3.15, indicating a larger amount of virus in NG samples across all gene targets tested. In gene targets where both sample types were negative, an assumed ΔCt difference was given as 0. Statistically significant differences in ΔΔCt values (p<0.05) are indicated with an asterisk (*). FIGS. 25A-B) Graphical representation includes data wherein at least 1 sample type was found to be positive for containing SARS-CoV-2, and negative Ct values were assigned a value of 40. FIG. 25C) NG sensitivity was 100%, whereas G sensitivity was 28.6% lower, i.e., NG sensitivity was 40% higher.

6.2: A Comparison Between Nasal Gargle and Anterior Nares Swabbing Sensitivity

[0943] Paired Nasal Gargle and anterior nares swabs (n=12) were compared by qPCR, with lower ΔCt values associated with Nasal Gargle over anterior nares swabbing (FIG. 26A). This result is of particular interest as anterior nares swabbing occurred prior to Nasal Gargle procurement, and without a wait period to permit SARS-CoV-2 containing material to re-establish, indicating that the Nasal Gargle was effective at sampling nasal regions that are generally inaccessible to anterior nares swabbing. Moreover, positive identification of SARS-CoV-2 occurred 22% more often in Nasal Gargle samples than in anterior nares swabs (FIG. 26B), demonstrating preferential sensitivity for the Nasal Gargle sampling technique.

[0944] Description of FIG. 26 (A Preliminary examination of Nasal Gargle Efficacy Over Anterior Nares Swabs): FIG. 26A) The average difference between Nasal Gargle and anterior nares swabs ΔCt values range from −0.45 to −0.727, indicating a larger amount of virus in Nasal Gargle samples across all gene targets tested. Graphical representation includes data wherein at least 1 sample type was found to be positive for containing SARS-CoV-2, and negative Ct values were assigned a value of 40. In gene targets where both sample types were negative, an assumed ΔCt difference was given as 0. FIG. 26B) Nasal Gargle sensitivity was 22% higher than anterior nares swab sensitivity.

6.3: Concentration of Nasal Gargle

[0945] To further increase the detection of SARS-CoV-2 in Nasal Gargle samples, Nasal Gargles were processed by a manual concentration device including two of the Polyester Filters described and characterized hereinabove in the description of Experiment 7. The subsequent and pairwise testing of Nasal Gargles before concentration, and material eluted from the concentration device by qPCR (n=4) demonstrated higher levels of viral material present after concentrating Nasal Gargles compared to unconcentrated Nasal Gargles. The average ΔΔCt values were −2.17 (RdRP/S), −2.17 (E), or −1.8975 (N) (FIG. 27B), corresponding to an estimated viral material increase of 3.857, 4.5, or 3.726-fold, respectively.

[0946] Paradoxically, in one patient, SARS-CoV-2 was detected weakly (38.03 Ct value) in a single gene target in unconcentrated Nasal Gargle, but in no gene targets after concentration. The high Ct value in one gene and negative readings for the other gene targets indicates that the viral load of the sample was at or near the limit of detection of the assay. It is likely that the swabbing of the Nasal Gargle prior to filtration, as described hereinabove in Section 5.1, selectively removed a significant portion of the virus, resulting in a sample that contained no or very little virus despite subsequent concentration.

[0947] Description of FIG. 27 (Additive Effects of the Concentration of Nasal Gargle Samples): FIG. 27A) Target gene ΔCt values of Nasal Gargle samples before concentration (NG=left endpoint) were paired with respective filters after NG concentration (Filter=right endpoint), after being normalized to a housekeeping gene tested in parallel. Filter ΔCt values tend to be less than unconcentrated NGΔCt values. FIG. 27B) The average difference between NG and filter ΔCt values range from −1.89 to −2.17, indicating a greater presence of virus due to concentration across all gene targets tested compared to unconcentrated NG. In gene targets where both sample types were negative, an assumed ΔCt difference was given as 0. FIGS. 27A-B) Graphical representation includes data wherein at least 1 sample type was found to be positive for containing SARS-CoV-2, and negative Ct values were assigned a value of 40.

Section 7: Discussion

[0948] The striking difference in the observed percentage of SARS-CoV-2 positive samples from Nasal Gargle compared to gargle or anterior nares swabs supports previous observations that viral replication does not always lead to the aggregation of viral material in the same physiological sites.sup.9, and suggests that Nasal Gargle is effective at sampling a greater number of virus containing regions, such as the nasopharynx, which are otherwise inaccessible to other self-sampling techniques. It therefore follows that utilizing the Nasal Gargle sampling method for current molecular and immunological devices would likely decrease the number of false negative tests. Specifically, our data supports the preferential usage of Nasal Gargle in PCR-based assays in order to prevent false negative test results, which can be damaging to public health and safety.

[0949] The preliminary clinical results demonstrate clear trends that have reached, or approach, statistical significance, of more desirable Ct values and sensitivity associated with Nasal Gargle sampling over that of gargle despite the small sample size. Furthermore, with similar results observed both by immunoassay and qPCR testing, we hypothesize that Nasal Gargle should provide a significant increase in sensitivity across a wide range of testing platforms. Impressively, Nasal Gargle outperformed gargle despite the patient sampling protocol requiring the procurement of anterior nares swabs, followed by gargle samples, prior to Nasal Gargle sampling, which might be expected to artificially decrease the amount of viral material in the oropharynx available for sampling by the subsequent Nasal Gargle. It is reasonable to assume that when paired samples are not being procured, such as during standard clinical testing, Nasal Gargle might demonstrate further increased sensitivity. Similarly, SARS-CoV-2 viral load peaks are associated with or before the presentation of symptoms, whereas severe symptoms generally develop one to two-weeks later, accounting for the observation that patients with severe symptoms present with a decreased viral load.sup.12. As our study population exclusively consisted of inpatients who were at an advanced stage of illness, it is therefore reasonable to assume that Nasal Gargle sampling might perform with further increased sensitivity during testing of the general population due to relatively higher levels of viral shedding. Moreover, we did not impose any restrictions on eating, drinking, brushing teeth, or smoking prior to sample collection, contrary to other studies evaluating the efficacy of gargle sampling.sup.1, which accentuates the increase in detection sensitivity observed with Nasal Gargle, as well as likely increasing sampling accessibility.sup.13.

[0950] Lastly, the observed correlation between concentration of Nasal Gargle and decreased Ct values is quite promising, especially when examined with the accumulative decrease of Ct values associated with the use of Nasal Gargle as the sample type. Furthermore, the increased sensitivity observed in Nasal Gargles, combined with an effective concentration protocol could overcome current difficulties in testing pooled gargle samples due to the dilution of SARS-CoV-2 containing material.sup.14,15, providing an effective medium for public screening.

Section 8: References

[0951] [1] Goldfarb, D. M., Tilley, P., Al-Rawahi, G. N., Srigley, J., Ford, G., Pedersen, H., Pabbi, A., Hannam-Clark, S., Charles, M. K., Dittrick, M., Gadkar, V. J., Pernica, J. M., & Hoang, L. (2020). Self-collected Saline Gargle Samples as an Alternative to Healthcare Worker Collected Nasopharyngeal Swabs for COVID-19 Diagnosis in Outpatients. MedRxiv, 2020.09.13.20188334. [0952] [2] Mittal, A., Gupta, A., Kumar, S., Surjit, M., Singh, B., Soneja, M., Soni, K. D., Khan, A. R., Singh, K., Naik, S., Kumar, A., Aggarwal, R., Nischal, N., Sinha, S., Trikha, A., & Wig, N. (2020). Gargle lavage as a viable alternative to swab for detection of SARS-CoV-2. Indian Journal of Medical Research, 152(1), 77. [0953] [3] Maricic, T., Nickel, O., Aximu-Petri, A., Essel, E., Gansauge, M., Kanis, P., Macak, D., Richter, J., Riesenberg, S., Bokelmann, L, Zeberg, H., Meyer, M., Borte, S., & Pääbo, S. (2020). A direct RT-qPCR approach to test large numbers of individuals for SARS-CoV-2. PloS One, 15(12), e0244824-e0244824. PubMed. [0954] [4] Stolte, D. (2020, October 26). Swish, Gargle, Repeat. UArizona Researcher Explores Mouth Rinse Test as Alternative to COVID-19 Nasal Swab. University of Arizona News. [0955] [5] Federal Drug Administration (FDA). (2020). Access Genetics, LLC-OraRisk COVID-19 RT-PCR EUA Summary—Updated Sep. 25, 2020. [0956] [6] Berenger, B. M., Fonseca, K., Schneider, A. R., Hu, J., & Zelyas, N. (2020). Sensitivity of Nasopharyngeal, Nasal and Throat Swab for the Detection of SARS-CoV-2. MedRxiv, 2020.05.05.20084889. [0957] [7] Little, B. P. (2021). False-Negative Nasopharyngeal Swabs and Positive Bronchoalveolar Lavage: Implications for Chest CT in Diagnosis of COVID-19 Pneumonia. Radiology, 204471. [0958] [8] Piras, A., Rizzo, D., Uzzau, S., De Riu, G., Rubino, S., & Bussu, F. (2020). Inappropriate Nasopharyngeal Sampling for SARS-CoV-2 Detection Is a Relevant Cause of False-Negative Reports. Otolaryngology—Head and Neck Surgery, 163(3), 459-461. [0959] [9] Kojima, N., Turner, F., Slepnev, V., Bacelar, A., Deming, L., Kodeboyina, S., & Klausner, J. (2020). Self-Collected Oral Fluid and Nasal Swabs Demonstrate Comparable Sensitivity to Clinician Collected Nasopharyngeal Swabs for Covid-19 Detection. MedRxiv, 2020.04.11.20062372. [0960] [10] Prince-Guerra, J. L., Almendares, O., Nolen, L. D., Gunn, J. K. L., Dale, A. P., Buono, S. A., Deutsch-Feldman, M., Suppiah, S., Hao, L., Zeng, Y., Stevens, V. A., Knipe, K., Pompey, J., Atherstone, C., Bui, D. P., Powell, T., Tamin, A., Harcourt, J. L., Shewmaker, P. L., Bower, W. A. (2021). Evaluation of Abbott BinaxNOW Rapid Antigen Test for SARS-CoV-2 Infection at Two Community-Based Testing Sites—Pima County, Ariz., Nov. 3-17, 2020. MMWR. Morbidity and Mortality Weekly Report, 70(3), 100-105. PubMed. [0961] [11] Agulló, V., Fernández-González, M., Ortiz de la Tabla, V., Gonzalo-Jiménez, N., García, J. A., Masiá, M., & Gutiérrez, F. (2020). Evaluation of the rapid antigen test Panbio COVID-19 in saliva and nasal swabs in a population-based point-of-care study. The Journal of Infection, S0163-4453(20)30768-4. PubMed. [0962] [12] Argyropoulos, K. V., Serrano, A., Hu, J., Black, M., Feng, X., Shen, G., Call, M., Kim, M. J., Lytle, A., Belovarac, B., Vougiouklakis, T., Lin, L. H., Moran, U., Heguy, A., Troxel, A., Snuderl, M., Osman, I., Cotzia, P., & Jour, G. (2020). Association of Initial Viral Load in Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Patients with Outcome and Symptoms. The American journal of pathology, 190(9), 1881-1887. [0963] [13] Kelly, A., & Steacy, L. (2021 Feb. 7). Gargle and spit COVID-19 test important tool to prevent “testing fatigue”: B.C. doctor. CityNews 1130. [0964] [14] Babady, N. E., McMillen, T., Jani, K., Viale, A., Robilotti, E. V., Aslam, A., Diver, M., Sokoli, D., Mason, G., Shah, M. K., Korenstein, D., & Kamboj, M. (2021). Performance of Severe Acute Respiratory Syndrome Coronavirus 2 Real-Time RT-PCR Tests on Oral Rinses and Saliva Samples. The Journal of Molecular Diagnostics: JMD, 23(1), 3-9. [0965] [15] Lüsebrink, J., Schildgen, V., & Schildgen, O. (2020). Pooling is an insufficient strategy to avoid healthcare staff to patient transmission of severe acute respiratory coronavirus virus 2 (SARS-CoV-2). Infection Control & Hospital Epidemiology, 1-3. Cambridge Core.

[0966] In some applications, any of the techniques described in the report of Experiment 8 above are used in combination with the other techniques described herein.

Experiment 9

[0967] Some of the techniques described hereinabove comprise intranasally dispensing nasal wash fluid into a nasal cavity of a subject; and thereafter, collecting a specimen sample by performing an anterior nares swab, without performing a mid-turbinate swab or a nasopharyngeal swab. Experiment 9 provides evidence of the efficacy of these techniques for collecting specimen samples for detection of SARS-CoV-2 using quantitative PCR (qPCR).

[0968] In summary, Experiment 9 found that spraying saline solution into the nasal cavity and gargling resulted in between a 3.4-fold (340%), and 5.6-fold (560%) increase in viral material after saline nasal spraying and gargling compared to before nasal spraying and gargling, depending on the gene target, after a nucleic acid extraction protocol consisting of including heating samples. Experiment 9 also found that spraying saline solution into the nasal cavity and gargling resulted in between a 2.7-fold (270%) and 8.7-fold (870%) increase in viral material after saline nasal spraying and gargling compared to before nasal spraying and gargling, depending on the gene target, after a robust magnetic bead based nucleic acid extraction protocol.

Detailed Report of Experiment 9: Benefits of Saline Nasal Spraying Before Anterior Nares Swabbing in SARS-CoV-2 Detection

Section 1: Materials and Methods

Section 1.1: Subject Sampling

[0969] Bilateral anterior nares swabs were performed by a healthcare professional on 18 subjects belonging to the health care service provider Leumit (Tel-Aviv, Israel), who had previously tested positive for SARS-CoV-2. The subjects then sprayed 0.1 mL of normal saline 3-5 times into each nostril (some while their heads were tilted back and others while their heads were in a neutral position), and then gargled 10 mL of normal saline for 10-15 seconds to produce a Nasal Gargle (hereinbelow, “NG”) specimen. Following spitting out of the NG sample, a second bilateral anterior nares swab was performed by the healthcare professional, while the subjects' heads were in a neutral position or tilted slightly back. Swabs were then placed into tubes containing 3 mL of Universal Transport Medium (UTM) (Biological Industries, Israel). Two subjects were excluded from the study due to an inability to gargle. For 3 subjects (Subjects 5-7), a bilateral mid-turbinate swab was performed by the healthcare professional prior to the production of the NG sample. Bilateral anterior nares swabbing was performed by inserting swabs into the nares and then removing the swabs without rotating the swabs against the inner wall of the nares. Samples were transported on ice to either the Leumit Health Services Central Laboratory (Or Yehuda, Israel), or to Hero Scientific Ltd. (Jerusalem, Israel) for processing. The clinical study was approved by the investigational review board (hereinbelow, “IRB”) at Assaf Harofeh (Shamir) Medical Center (Tel Aviv, Israel).

[0970] It is important to note that the anterior nares swab sample collection was performed by inserting the swab about 1.5 cm into the nostril and spinning the swab vertically as is typically performed for a mid-turbinate swab sample collection. All anterior nares swab sample collection procedures performed for all other experiments described herein (Experiments 10, 11, 12, 13, 14, and 15) were performed by inserting the swab about 1.5 cm into the nostril and rubbing the swab in a circular motion so as to abrasively rub against the wall of the anterior naris. The inventors hypothesize that this experiment may have been negatively impacted by this non-ideal anterior nares swab sample collection procedure. In particular, the anterior nares swab sample performed after saline nasal administration may have been unable to properly collect the saline dripping down the wall of the anterior naris and thus not yield the benefits of saline nasal administration in increasing the viral load collected. Conversely, the inventor hypothesize that other experiments described herein (Experiments 10, 11, 12, 13, 14, and 15) are more adequate representations of the benefit of anterior nares swab sampling after saline nasal administration.

Section 1.2: Molecular Testing

[0971] Section 1.2.1: qPCR Following Heat Extraction

[0972] Samples from 14 of the 16 subjects were processed by quantitative reverse transcription polymerase chain reaction (hereinbelow, “qPCR”) at Hero Scientific Ltd. (Jerusalem, Israel). Samples were first inactivated by boiling at 70° C. for 45 minutes in a heat block with aluminum beads. All samples were processed using the primers and probe sequences from the Center for Disease Control's (hereinbelow, “CDC”) diagnostic panel provided in the 2019-nCoV RUO Kit (CAT #10006713, IDT, Belgium) on an MX3000P (Stratagene, USA) qPCR thermocycler. The reaction mixture for samples from 3 of the 14 subjects (Subjects 5-7) were prepared using the GE50 Xpert One-Step Fast Probe (GRiSP Research Solutions, Portugal), and contained 10 microliters of Fast qPCR Mastermix (GRiSP Research Solutions, Portugal), 0.5 microliters of RTase Mix (GRiSP Research Solutions, Portugal), 2 microliters of primer mix (CAT #10006713, IDT, Belgium), 4 microliters of inactivated sample, and 3.5 microliters of Molecular Grade Water (Biological Industries, Israel). Negative template controls (hereinbelow, “NTCs”) were performed by testing 4 microliters of Molecular Grade Water (Biological Industries, Israel) instead of the samples for all primer/probe mixtures tested. Similarly, positive controls were performed by testing 1 microliter of positive control plasmid, 2019-nCoV_N_Positive Control (CAT #10006625, IDT, Belgium), and 3 microliters of Molecular Grade Water (Biological Industries, Israel) instead of the samples for all primer/probe mixtures tested. The thermal profile conditions were 45° C. for 15 minutes, 95° C. for 3 minutes, and then 40 cycles of 95° C. for 5 seconds and 58° C. for 30 seconds. Prior to use of the GE50 Xpert One-Step Fast Probe, 2 microliters of 100×ROX was added to the Fast qPCR Mastermix (Probe) as per the manufacturer's instructions, and cycle threshold (hereinbelow, “Ct”) values were normalized to ROX. (As is known in the quantitative PCR art, the Ct value is defined as the number of cycles of amplification (using PCR) required for the fluorescence of a PCR product (i.e., the target/amplicon) to be detected crossing a threshold that is above the background signal. Relatively lower Ct values indicate higher initial viral levels.)

[0973] Samples from 12 of the 14 subjects (including Subjects 1-4 and 8-11) were processed using the PrimeDirect Probe RT-qPCR Mix (Takara Bio inc., Japan). Reaction mixtures contained 12.5 microliters of the PrimeDirect Probe RT-qPCR Mix (Takara Bio inc., Japan), 4 microliters of sample, 2 microliters of the primer/probe mix (IDT, Belgium), and 6.5 microliters of Molecular Grade Water (Biological Industries, Israel). NTCs were performed for each qPCR run by testing 4 microliters of Molecular Grade Water instead of the samples for all primer/probe mixtures tested. Similarly, positive controls were performed by testing 1 microliter of positive control plasmid, 2019-nCoV_N_Positive Control (CAT #10006625, IDT, Belgium), and 3 microliters of Molecular Grade Water (Biological Industries, Israel) instead of the samples for all primer/probe mixtures tested. The thermal profile conditions were 90° C. for 3 minutes, 60° C. for 5 minutes, and then 40 cycles of 95° C. for 5 seconds and 60° C. for 30 seconds.

Section 1.2.2: qPCR Following Magnetic Bead Extraction

[0974] Samples from 2 of the 16 subjects (Subjects 12 and 13) were processed by qPCR at the Leumit Health Services Central Laboratory (Or Yehuda, Israel) using the Allplex 2019 n-CoV Assay (Seegene, Seoul, South Korea), following manufacturer's instructions, on a CFX96 Real-Time System (BIO RAD, USA). Prior to qPCR, samples were extracted using the STARMag 96×4 Viral DNA/RNA 200 C Kit (Seegene, Seoul, South Korea) on a MICROLAB STAR.sup.let (Hamilton, Switzerland) automated liquid handling platform.

Section 2: Results and Discussion

[0975] Out of the 16 subjects, 13 (n=13) were found to be positive for SARS-CoV-2 on at least one gene target for either of the paired anterior nares swabs. Average Ct value differences between (a) anterior nares swabs sampled before saline nasal spraying and (b) anterior nares swabs samples after saline nasal spraying and gargling for SARS-CoV-2 gene targets, after a nucleic acid extraction protocol including heating samples, ranged from −1.75 to −2.5, corresponding to between a 3.4-fold (340%), and 5.6-fold (560%) increase in viral material after saline nasal spraying and gargling compared to before nasal spraying and gargling, depending on the gene target (Table 7.1). Similarly, average ΔCt value differences between (a) anterior nares swabs sampled before saline nasal spraying and (b) anterior nares swabs samples after saline nasal spraying and gargling for SARS-CoV-2 gene targets, after a robust magnetic bead based nucleic acid extraction protocol, ranged from −1.45 to −3.12, corresponding to between a 2.7-fold (270%) and 8.7-fold (870%) increase in viral material after saline nasal spraying and gargling compared to before nasal spraying and gargling, depending on the gene target (Table 7.2). This phenomenon is likely due to the saline introduced during NG sampling dripping from within the nasal cavity back into the anterior nares, increasing the amount of viral material available for subsequent swabbing. It is therefore expected that performing nasal spray prior to anterior nares swabbing would lead to an increase in sensitivity in any downstream molecular or immunological tests performed using the swabbed specimen samples. Interestingly, similar results were observed across different RNA extraction and qPCR protocols, indicating that a combined nasal spray and anterior nares swab sampling methodology would be effective at increasing diagnostic sensitivity even when used in conjunction with devices where less-robust nucleic acid extraction methods are employed.

TABLE-US-00007 TABLE 7.1 A Comparison Between Anterior Nares Swabbing Before And After Nasal Gargling Following Heat Extraction 1.sup.st 2.sup.nd Anterior Anterior Nares Nares Average Ct Gene Subject Swab Ct Swab Ct Ct Value Value Target # Value.sup.i Value.sup.i Difference.sup.ii Difference.sup.ii N1  1 32.73 34.84 2.11 −1.7536364  2 34.91 40 5.09  3 39.05 31.84 −7.21  4 40 40 0  5.sup.iii 31.7 24.15 −7.55  6.sup.iii 17.95 26.54 8.59  7.sup.iii 31.27 24.14 −7.13  8 39.52 38.2 −1.32  9 27.62 27.92 0.3 10 40 34.23 −5.77 11 31.98 25.58 −6.4 N2  1 40 40 0 −2.4954545  2 40 40 0  3 40 31.72 −7.01  4 40 39.23 −0.77  5.sup.iii 36.58 25.06 −11.52  6.sup.iii 19.69 28.81 9.12  7.sup.iii 34.53 26.84 −7.69  8 40 38.68 −1.32  9 32.81 29.32 −3.49 10 40 38.62 −1.38 11 33.69 30.3 −3.39 .sup.iNegative Ct values, corresponding to a value of ≥40, were assigned an actual value of 40. .sup.iiIn instances where both swabs were negative, the average Ct value difference was assumed to bo 0. .sup.iiiThe NTC accompanying the tests for Subjects 5-7 had evidence of human DNA contamination. However, if human DNA contamination were to occur in subject samples, it would be unlikely to affect results for SARS-CoV-2 specific gene targets, and so the reults were included, but not normalized to the reference gene, RNase P.

TABLE-US-00008 TABLE 7.2 A Comparison Between Anterior Nares Swabbing before and after Nasal Gargling Following Magnetic Bead Extraction 1.sup.st 2.sup.nd Normalized Normalized Anterior Anterior 1.sup.st Anterior 2.sup.nd Anterior Nares Nares Nares Nares Normalized Average Gene Subject Swab Ct Swab Ct Swab ΔCt Swab ΔCt ΔCt Value ΔCt Value Target # Value Value Value.sup.i Value.sup.i Difference.sup.ii Difference E 12 24.83 26.72 −1.4 0.23 1.63 −1.73 13 35.04 29.57 8.5 3.41 −5.09 RdRP/S 12 26.93 28.55 0.7 2.06 1.36 −3.12 13 39.19 31.21 12.65 5.05 −7.6 N 12 27.77 29.42 1.54 2.93 1.39 −1.45 13 37.16 32.49 10.62 6.33 −4.29 .sup.iNormalized ΔCt value is calculated by subtracting the Ct value of a reference gene from the Ct value of the target gene. .sup.iiNormalized ΔCt value difference is calculated by subtracting the normalized ΔCt value of the anterior nares swab performed prior to the NG (1.sup.st) from the normalized ΔCt value of the anterior nares swab performed after the NG (2.sup.nd).

[0976] In some applications, any of the techniques described in the report of Experiment 9 above are used in combination with the other techniques described herein.

Experiment 10

[0977] Some of the techniques described hereinabove comprise intranasally dispensing nasal wash fluid into a nasal cavity of a subject; and thereafter, collecting a specimen sample by performing an anterior nares swab, without performing a mid-turbinate swab or a nasopharyngeal swab. Optionally, the method further comprises testing for the presence of a particulate, such as a virus (e.g., SARS-CoV-2), for example using PCR. Experiment 10 provides evidence of the efficacy of these techniques for collecting specimen samples for detection of SAILS-CoV-2 using quantitative PCR (qPCR).

[0978] In summary, Experiment 10 found that eleven out of 15 subjects tested (73.3%) had lower normalized cycle threshold (Ct) values in samples from anterior nares swabs performed after saline nasal spraying and gargling than from anterior nares swabs performed before saline nasal spraying and gargling. Furthermore, 7 out of the 15 subjects (46.7%) were found to have lower normalized Ct values in samples from anterior nares swabs performed after saline nasal spraying and gargling than in mid-turbinate swabs. In cases where the mid-turbinate swab outperformed the anterior nares swabs sampled after saline nasal spraying and gargling, anterior nares swabs sampled after saline nasal spraying and gargling were still within 1 ΔCt value of the mid-turbinate swab in 5 out 8 subjects (62.5%).

Detailed Report of Experiment 10: Benefits of Saline Nasal Spraying Prior to Anterior Nares Swabbing in SARS-CoV-2 Detection on the Cobas 6800 System

Section 1: Materials and Methods

Section 1.1: Subject Sampling

[0979] Bilateral anterior nares swabbing was performed by a healthcare professional on 15 subjects belonging to the health care service provider Leumit (Tel-Aviv, Israel), who had previously tested positive for SARS-CoV-2. A bilateral mid-turbinate swab was then performed by the healthcare professional. The subjects then sprayed 0.1 mL of normal saline 3-5 times into each nostril, and then gargled 10 mL of normal saline for 10-15 seconds to produce a Nasal Gargle (hereinbelow, “NG”) specimen. Following spitting out of the NG sample, a second bilateral anterior nares swab was performed by the healthcare professional. Swabs were then placed into tubes containing 3 mL of Universal Transport Medium (UTM) (Biological Industries, Israel). Samples were transported on ice to the Leumit Health Services Central Laboratory (Or Yehuda, Israel), for processing. The clinical study was approved by the investigational review board (IRB) at Assaf Harofeh (Shamir) Medical Center (Tel Aviv, Israel). All swabbing was performed with flocked swabs (3BY Ltd., Industrial Park Tefen, Ref #: 3by-001).

Section 1.2: Molecular Testing

[0980] RNA extraction and qPCR were performed using the Cobas SARS-CoV-2 Test (Roche, Switzerland) on a Cobas 6800 system (Roche, Switzerland), in accordance with manufacturer instructions.

Section 2: Results and Discussion

[0981] All 15 subjects were positive for both ORF1 a/b and E gene targets (Table 8). (As is known in the quantitative PCR art, the Ct value is defined as the number of cycles of amplification (using PCR) required for the fluorescence of a PCR product (i.e., the target/amplicon) to be detected crossing a threshold that is above the background signal. Relatively lower Ct values indicate higher initial viral levels.)

[0982] Fourteen out of the 15 subjects (93.3%) were found to have lower Ct values, indicative of higher viral levels, for both gene targets in samples from anterior nares swabs performed after saline nasal spraying and gargling than from anterior nares swabs performed before saline nasal spraying and gargling. Furthermore, 7 out of the 15 subjects (46.7%) were found to have lower Ct values, indicative of higher viral levels, across both gene targets in samples from anterior nares swabs performed after saline nasal spraying and gargling than in mid-turbinate swabs. In cases where the mid-turbinate swab outperformed the anterior nares swabs sampled after saline nasal spraying and gargling on at least one gene target, anterior nares swabs sampled after saline nasal spraying and gargling were still within 1 Ct value of the mid-turbinate swab in 4 out 8 subjects (62.5%), with 2 of the subjects outperforming mid-turbinate swabs on the second target.

[0983] On average, Ct values for anterior nares swabs performed after saline nasal spraying decreased by 2.35 for the ORF1 a/b gene and by 2.70 for the E gene compared to Ct values for anterior nares swabs performed before saline nasal spraying (Table 9). This decrease was statistically significant (ORF1 a/b gene, p=0.00014; E gene, p=0.00125) (Student's t-test).

[0984] This data suggest that dispensing nasal spray and gargling before anterior nares swabbing increases collected viral material when compared to anterior nares swabs without a prior nasal spray and gargle, and in some cases, when compared to mid-turbinate swabs. It is therefore expected that administering a nasal spray prior to anterior nares swabbing as a sampling method would result in an increase in molecular and immunological testing sensitivity.

TABLE-US-00009 TABLE 8 Ct value results Subject Sample.sup.1 ORF1 a/b gene E gene IC.sup.ii 1 AN1 20.38 20.45 32.91 MT 19.53 19.66 33.12 AN2 19.42 19.66 32.9 2 AN1 34.05 39.74 32.83 MT 30.8 31.6 33.04 AN2 29.78 30.51 32.81 3 AN1 24.36 24.14 32.33 MT 23.27 23.27 32.68 AN2 23.32 23.14 32.59 4 AN1 32.73 33.79 33.13 MT 28.71 29.19 32.75 AN2 30.54 31.35 32.77 5 AN1 27.04 27.1 32.27 MT 27.51 27.7 32.68 AN2 28.11 28.51 32.59 6 AN1 26.62 26.87 32.68 MT 25.24 25.46 32.53 AN2 23.87 24.07 32.42 7 AN1 29.89 30.44 32.89 MT 24.82 24.76 32.35 AN2 26.89 27.24 32.55 AN1 28.19 28.54 32.66 8 MT 29.52 30.1 32.76 AN2 26.43 26.64 32.23 9 AN1 21.24 21.48 32.59 MT 20.98 21.29 32.91 AN2 19.09 19.1 33.5 10 AN1 30.05 30.69 33.11 MT 30.17 30.99 33.11 AN2 29.12 29.96 32.72 11 AN1 34.26 35.8 32.82 MT 29.5 30.48 33.38 AN2 29.07 30.01 32.73 12 AN1 30.62 31.18 32.89 MT 23.37 23.59 32.7 AN2 25.17 25.7 34.19 13 AN1 30.04 30.43 32.94 MT 27.27 27.57 32.61 AN2 27.67 27.97 32.37 14 AN1 27.84 28.34 32.7 MT 27.03 27.64 32.81 AN2 24.4 24.88 32.76 15 AN1 35.18 36.79 32.89 MT 33.02 34.4 32.82 AN2 34.33 36.49 32.73 .sup.iAN1 = anterior nares swab performed prior to saline nasal spraying and gargling, MT = mid-turbinate swab, AN2 = anterior nares swab performed after saline nasal spraying and gargling .sup.iiIC = internal control

TABLE-US-00010 TABLE 9 Average Ct values and Average Difference between AN1 and AN2 AN1 AN2 Delta: AN2 − AN1 ORF1 a/b ORF1 a/b ORF1 a/b Subject gene E gene gene E gene gene E gene 1 20.38 20.45 19.42 19.66 −0.96 −0.79 2 34.05 39.74 29.78 30.51 −4.27 −9.23 3 24.36 24.14 23.32 23.14 −1.04 −1 4 32.73 33.79 30.54 31.35 −2.19 −2.44 5 27.04 27.1 28.11 28.51 1.07 1.41 6 26.62 26.87 23.87 24.07 −2.75 −2.8 7 29.89 30.44 26.89 27.24 −3 −3.2 8 28.19 28.54 26.43 26.64 −1.76 −1.9 9 21.24 21.48 19.09 19.1 −2.15 −2.38 10 30.05 30.69 29.12 29.96 −0.93 −0.73 11 34.26 35.8 29.07 30.01 −5.19 −5.79 12 30.62 31.18 25.17 25.7 −5.45 −5.48 13 30.04 30.43 27.67 27.97 −2.37 −2.46 14 27.84 28.34 24.4 24.88 −3.44 −3.46 15 35.18 36.79 34.33 36.49 −0.85 −0.3 Average 28.83 29.72 26.48 27.01 −2.35 −2.70

[0985] In some applications, any of the techniques described in the report of Experiment 10 above are used in combination with the other techniques described herein. It is noted that although Experiment 10, as reported above, included mid-turbinate swabbing of the subjects, this mid-turbinate swabbing served for comparison with anterior nares swabbing. Some techniques of the present invention include performing only an anterior nares swab, without performing a mid-turbinate swab.

Experiment 11

[0986] Some of the techniques described hereinabove comprise intranasally dispensing nasal wash fluid into a nasal cavity of a subject; and thereafter, collecting a specimen sample by performing an anterior nares swab, without performing a mid-turbinate swab or a nasopharyngeal swab. Optionally, the method further comprises testing for the presence of a particulate, such as a virus (e.g., SARS-CoV-2), for example using an immunoassay. Experiment 11 provides evidence of the efficacy of these techniques for collecting specimen samples for detection of SARS-CoV-2 using lateral flow immunoassay strips.

[0987] In summary, Experiment 11 found that two of the four subjects tested positive for SARS-CoV-2 antigens both on the mid-turbinate swab and on the anterior flares swab performed after saline nasal spraying, but negative on the anterior nares swab performed before saline nasal spraying, indicating that saline nasal spraying can increase anterior nares swab sensitivity with immunoassays. The remaining two of the four subjects tested negative for SARS-CoV-2 antigens on all sample types (anterior nares swabbing before saline nasal spraying, mid-turbinate swabbing, and anterior nares swabbing after saline nasal spraying).

Detailed Report of Experiment 11: Benefits of Saline Nasal Spraying Prior to Anterior Nares Swabbing in SARS-CoV-2 Detection by Immunoassay

Section 1: Materials and Methods

Section 1.1: Subject Sampling

[0988] Bilateral anterior nares swabbing was performed by a healthcare professional on 20 subjects belonging to the health care service provider Leumit (Tel-Aviv, Israel), who had previously tested positive for SARS-CoV-2. The subjects then sprayed 0.1 mL of normal saline 3-5 times into each nostril, and then gargled 10 mL of normal saline for 10-15 seconds to produce a Nasal Gargle specimen. Following spitting out of the Nasal Gargle sample, a second bilateral anterior nares swab was performed by the healthcare professional. Lastly, a bilateral mid-turbinate swab was performed by the healthcare professional. Dry swab samples were transported in 15 mL conical tubes (LabCon, USA, Ref #: LC 3136-345-008-9) on ice to Hero Scientific Ltd. (Jerusalem, Israel) for processing. All swabbing was performed with flocked swabs (3BY Ltd., Industrial Park Tefen, Ref #: 3by-001). The clinical study was approved by the investigational review board (IRB) at Assaf Harofeh (Shamir) Medical Center (Tel Aviv, Israel).

Section 1.2: Immunological Testing

[0989] Immunoassays were performed using either the BD Veritor™ system for Rapid Detection of SARS-CoV-2 (Becton Dickinson and Company, Maryland, USA, Ref: 256082), or the COVID-19 Antigen Rapid Test Cassette (Beijing Kewei Clinical Diagnostic Reagent Inc., Beijing, China, Ref: 601450) in accordance with manufacturer instructions. The BD Veritor™ system for Rapid Detection of SARS-CoV-2 is a chromatographic digital immunoassay intended for the direct and qualitative detection of SARS-CoV-2 nucleocapsid antigens in nasal swabs, and the COVID-19 Antigen Rapid Test Cassette is a solid phase immunochromatographic assay for the in vitro qualitative detection of SARS-CoV-2 antigen in nasopharyngeal, nasal, and throat swab specimens. Due to the limited availability of the BD Veritor™ system for Rapid Detection of SARS-CoV-2, in 4 subjects the anterior nares performed prior to nasal spraying and gargling was tested on the COVID-19 Antigen Rapid Test Cassette, with all other sample types tested on the BD Veritor™ system for Rapid Detection of SARS-CoV-2. In order to determine semi-quantitative differences in viral load between sample types, band intensity was scored on a scale of 0-3, with 0 corresponding to a negative test.

Section 2: Results and Discussion

[0990] 7 subjects tested negative and 4 subjects tested positive for SARS-CoV-2 antigens on all sample types (anterior nares swabbing before saline nasal spraying and gargling, mid-turbinate swabbing, and anterior nares swabbing after saline nasal spraying and gargling).

[0991] 6 out of the 9 subjects with discordant results (67%) tested positive for SARS-CoV-2 antigens both on the mid-turbinate swab and on the anterior nares swab performed after saline nasal spraying and gargling, but negative on the anterior nares swab performed before saline nasal spraying and gargling, indicating that saline nasal spraying increases anterior nares swab efficacy in viral material sampling and sensitivity with immunoassays. In a like manner, 1 out of the 9 subjects with discordant results tested clearly positive for SARS-CoV-2 antigens both on the mid-turbinate swab and on the anterior nares swab performed after saline nasal spraying and gargling; however, on the anterior nares swab prior to saline nasal spraying and gargling, it was unclear whether or not there was a faint band corresponding to the presence of SARS-CoV-2 antigens. Regarding the remaining 2 subjects with discordant results, 1 subject tested positive for SARS-CoV-2 antigens on the mid-turbinate swab, but negative on both anterior nares swabs, and 1 subject tested positive for SARS-CoV-2 antigens on the anterior nares swab performed after saline nasal spraying and gargling, but negative on both the mid-turbinate and anterior nares swab performed prior to nasal spraying and gargling. Similarly, positive band intensity for SARS-CoV-2 antigens in anterior nares swab samples performed after saline nasal spraying and gargling was equal to that of the mid-turbinate swab samples for 2 out of the 15 subjects (13.3%) that presented a positive on both the anterior nares swab performed after saline nasal spraying and gargling and the mid-turbinate swab. Moreover, positive band intensity for SARS-CoV-2 antigens was higher in anterior nares swab samples performed after saline nasal spraying and gargling than that of the mid-turbinate swab samples for 3 out of the 15 subjects (20%) that presented a positive on both the anterior nares swab performed after saline nasal spraying and gargling and the mid-turbinate swab, indicating that by coupling nasal spray with anterior nares swabbing, anterior nares swabs in some cases collect more viral material than mid-turbinate swabs.

[0992] In some applications, any of the techniques described in the report of Experiment 11 above are used in combination with the other techniques described herein. It is noted that although Experiment 11, as reported above, included mid-turbinate swabbing of the subjects, this mid-turbinate swabbing served for comparison with anterior nares swabbing. Some techniques of the present invention include performing only an anterior nares swab, without performing a mid-turbinate swab.

Experiment 12

[0993] Some of the techniques described hereinabove comprise intranasally dispensing nasal wash fluid into a nasal cavity of a subject; and thereafter, collecting a specimen sample by performing an anterior nares swab, without performing a mid-turbinate swab or a nasopharyngeal swab. Optionally, the method further comprises testing for the presence of a particulate, such as a virus (e.g., SARS-CoV-2), for example using PCR. Experiment 12 provides evidence of the efficacy of these techniques for collecting specimen samples for detection of SARS-CoV-2 using quantitative PCR (qPCR).

Detailed Report of Experiment 12: Benefits of Saline Nasal Spraying Before Anterior Nares Swabbing in SARS-CoV-2 Detection

Section 1: Materials and Methods

Section 1.1: Subject Sampling

[0994] Three bilateral anterior nares swab samplings were performed approximately one minute apart by a healthcare professional on 12 subjects belonging to the health care service provider Leumit (Tel-Aviv, Israel), who had previously tested positive for SARS-CoV-2. The subjects then sprayed 0.1 mL of normal saline 3-5 times into each nostril and gargled 10 mL of normal saline. Then a fourth bilateral anterior nares swab was performed by the healthcare professional. Bilateral anterior nares swabbing was performed by inserting swabs into the nares and then removing the swabs without rotating the swabs against the inner wall of the nares. Swabs were then placed into tubes containing 3 mL of Universal Transport Medium (UTM) (Biological Industries, Israel). Samples were transported on ice to the Leumit Health Services Central Laboratory (Or Yehuda, Israel), for processing. The clinical study was approved by the investigational review board (IRB) at Assaf Harofeh (Shamir) Medical Center (Tel Aviv, Israel). All swabbing was performed with flocked swabs (3BY Ltd., Industrial Park Tefen, Israel, Ref: 3by-001).

Section 1.2: Molecular Testing

[0995] Samples were processed by qPCR at the Leumit Health Services Central Laboratory (Or Yehuda, Israel) using the Allplex 2019 n-CoV Assay (Seegene, Seoul, South Korea), following manufacturer's instructions, on a CFX96 Real-Time System (BIO RAD, USA). Prior to qPCR, samples were extracted using the STARMag 96×4 Viral DNA/RNA 200 C Kit (Seegene, Seoul, South Korea) on a MICROLAB STARlet (Hamilton, Switzerland) automated liquid handling platform. Statistical analysis was performed using the Wilcoxon signed rank test to determine p-values.

Section 2: Results and Discussion

[0996] Lower Ct values were observed in anterior nares swabs performed after nasal spraying and gargling than before nasal spraying and gargling on average across all SARS-CoV-2 gene targets (Tables 10.1, 10.2), indicative of higher viral levels after nasal spraying and gargling than before nasal spraying and gargling independent of possible contributions from a mid-turbinate swab.

[0997] Average Ct values of the fourth anterior nares swab, which were preceded by nasal spraying and gargling, were lower across all gene targets than that of the other paired anterior nares swabs performed prior to nasal spraying and gargling (Table 10.2). The reversal of the observed aforementioned trend highlights the benefit of performing nasal spraying and gargling prior to anterior nares swabbing in SARS-CoV-2 detection, specifically in light of comparisons between the third anterior nares swab performed prior to nasal spraying and gargling and the fourth anterior nares swab performed following nasal spraying and gargling. The difference between the averages of the third nares swab performed prior to nasal spraying and gargling and the fourth anterior nares swab performed following nasal spraying and gargling reached statistical significance across all gene targets tested (Table 10.3) despite the relatively small sample size.

TABLE-US-00011 TABLE 10.1 Ct calue results C(t) values for each Gene Target.sup.ψ Subject # Sample.sup.i E RdRP/S N IC.sup.ii 1 AN1 40.0 40.0 40.0 25.6 AN2 33.7 36.0 36.0 27.1 AN3 40.0 40.0 40.0 27.4 AN4 (after NG) 34.7 35.1 37.8 25.8 2 AN1 25.6 27.5 37.8 26.7 AN2 28.8 31.5 27.8 26.9 AN3 26.2 28.7 31.1 25.7 AN4 (after NG) 24.8 26.8 26.9 25.8 3 AN1 25.0 26.9 27.2 25.7 AN2 24.3 26.1 26.5 25.4 AN3 23.0 25.0 24.4 24.8 AN4 (after NG) 20.1 21.7 22.3 27.1 4 AN1 40.0 40.0 40.0 40.0 AN2 40.0 40.0 40.0 40.0 AN3 40.0 40.0 40.0 40.0 AN4 (after NG) 40.0 40.0 40.0 40.0 5 AN1 36.4 40.0 39.5 26.7 AN2 40.0 40.0 36.9 25.5 AN3 40.0 40.0 40.0 26.7 AN4 (after NG) 40.0 40.0 40.0 25.2 6 AN1 28.4 30.5 31.3 26.7 AN2 29.2 30.9 31.7 25.5 AN3 30.4 32.6 33.0 26.7 AN4 (after NG) 28.0 29.4 30.4 25.2 7 AN1 40.0 40.0 37.9 25.7 AN2 40.0 36.7 40.0 25.6 AN3 40.0 40.0 40.0 26.9 AN4 (after NG) 40.0 40.0 40.0 27.1 8 AN1 40.0 40.0 40.0 25.6 AN2 40.0 40.0 40.0 27.1 AN3 40.0 40.0 40.0 26.9 AN4 (after NG) 40.0 37.1 40.0 25.4 9 AN1 40.0 40.0 40.0 27.2 AN2 40.0 40.0 40.0 27.3 AN3 40.0 40.0 40.0 27.1 AN4 (after NG) 40.0 39.2 40.0 27.3 10 AN1 40.0 40.0 40.0 40.0 AN2 40.0 40.0 40.0 40.0 AN3 40.0 40.0 40.0 40.0 AN4 (after NG) 40.0 40.0 40.0 40.0 11 AN1 32.3 34.2 33.7 25.8 AN2 32.8 34.5 33.7 25.5 AN3 31.9 33.3 34.4 26.9 AN4 (after NG) 28.3 29.6 30.3 26.0 12 AN1 24.8 26.6 26.7 25.4 AN2 29.6 31.1 31.6 25.5 AN3 26.4 28.0 28.8 25.7 AN4 (after NG) 25.9 27.8 28.3 25.8 .sup.iAN1, AN2, AN3 = Three anterior nares swabs performed in succession prior to nasal spraying and gargling (NG); AN4 = Anterior nares swab performed after NG .sup.iiIC = Internal control used in molecular assay .sup.ψNegative Ct values are expressed as 40

TABLE-US-00012 TABLE 10.2 Ct value aver uses.sup.iii C(t) value averages for each Gene Target Sample E RdRP/S N AN1 31.56 33.21 34.27 AN2 32.30 33.36 33.04 AN3 32.24 33.45 33.95 AN4 (after NG) 30.21 31.29 31.99 .sup.iiiFor the purpose of making a meaningful comparison between swab samples, average Ct values exclude subjects with less than two positive gene targets across all samples (excluded: subject 4, subject 8, subject 9, and subject 10)

TABLE-US-00013 TABLE 10.3 Statistical Significance: p-values.sup.iv p-values for each Gene Target Paired Samples E RdRP/S N AN1-AN2 0.447 0.866 0.554 AN1-AN3 0.344 0.590 0.933 AN1-AN4 0.272 0/142 0.195 AN2-AN3 0.675 0/800 0.272 AN2-AN4 0.059 0.076 0.272 AN3-AN4 0.036* 0.036* 0.036* .sup.ivThe Wilcoxon signed rank test was used to determine p-values. For the purpose of making a meaningful comparison between swab samples, statistical tests exclued subjects with less than two positive gene targets across all samples (excluded: subject 4, subject 8, subject 9, and subject 10) *Indicates statistically significant values

[0998] In some applications, any of the techniques described in the report of Experiment 12 above are used in combination with the other techniques described herein.

Experiment 13

[0999] Some of the techniques described hereinabove comprise intranasally dispensing nasal wash fluid into a nasal cavity of a subject; and thereafter, collecting a specimen sample by performing an anterior nares swab, without performing a mid-turbinate swab or a nasopharyngeal swab. Optionally, the method further comprises testing for the presence of a particulate, such as a virus (e.g., SARS-CoV-2), for example using an immunoassay. Experiment 13 provides evidence of the efficacy of these techniques for collecting specimen samples for detection of SARS-CoV-2 using lateral flow immunoassay strips.

[1000] In summary, Experiment 13 found that, in a single subject, the SARS-CoV-2 antigen band intensity for an anterior nares swab performed following nasal saline spraying was greater than that of both an anterior flares swab performed prior to nasal saline spraying and a mid-turbinate swab, indicating a greater quantity of virus recovered on an anterior nares swabs performed following nasal saline spraying than on anterior flares swab performed prior to nasal saline spraying and a mid-turbinate swab.

Detailed Report of Experiment 13: Further Evaluation of the Benefits of Saline Nasal Spraying without Gargling Prior to Anterior Nares Swabbing in SARS-CoV-2 Detection by Immunoassay

Section 1: Materials and Methods

Section 1.1: Subject Sampling

[1001] Bilateral anterior nares swabbing was performed by a healthcare professional on 7 subjects belonging to the health care service provider Leumit (Tel-Aviv, Israel), who had previously tested positive for SARS-CoV-2. The subjects then sprayed 0.1 mL of normal saline 3-5 times into each nostril with their heads in a neutral position or slightly tilted back. Subjects then significantly tilted their heads backwards by extending their necks as far as comfortably possible and maintained this head positioning for 5-10 seconds, similar to the conditions that normally occur during gargling. The subjects then returned their heads to a neutral position and a second bilateral anterior nares swabbing was performed by the healthcare professional. Lastly, bilateral mid-turbinate swabbing was performed by the healthcare professional. Dry swab samples were transported in 15 mL conical tubes (LabCon, USA, Ref #: LC 3136-345-008-9) on ice to Hero Scientific Ltd. (Jerusalem, Israel) for processing. All swabbing was performed with flocked swabs (3BY Ltd., Industrial Park Tefen, Ref #: 3by-001). The clinical study was approved by the investigational review board (IRB) at Assaf Harofeh (Shamir) Medical Center (Tel Aviv, Israel).

Section 1.2: Immunological Testing

[1002] Immunoassays were performed on all sample types using the BD Veritor™ system for Rapid Detection of SARS-CoV-2 (Becton Dickinson and Company, Maryland, USA, Ref: 256082), in accordance with manufacturer instructions. The BD Veritor™ system for Rapid Detection of SARS-CoV-2 is a chromatographic digital immunoassay intended for the direct and qualitative detection of SARS-CoV-2 nucleocapsid antigens in nasal swabs. In order to determine semi-quantitative differences in viral load between sample types, band intensity was scored on a scale of 0-3, with 0 corresponding to a negative test.

Section 2: Results and Discussion

[1003] Six of the seven subject samples were close to their 10-day quarantine release date, or had already been released, and unsurprisingly tested negative on all immunoassays performed. The single remaining subject who had tested positive for SARS-CoV-2 by qPCR 4 days prior to sample collection tested positive for the presence of SARS-CoV-2 antigen on all sample types (anterior nares swab prior to nasal saline spraying, anterior nares swab following nasal saline spraying, and the mid-turbinate swab) by immunoassay. The SARS-CoV-2 antigen band intensity for the anterior nares swab performed following nasal saline spraying (score: 3) was greater than that of both the anterior nares swab performed prior to nasal saline spraying (score: 2) and the mid-turbinate swab (score: 2.5), indicating a greater quantity of virus recovered on an anterior nares swab performed following nasal saline spraying than on an anterior nares swab performed prior to nasal saline spraying and a mid-turbinate swab.

[1004] This observation supports the fact that coupling nasal saline spraying to anterior nares swabbing increases immunoassay testing sensitivity. As subjects did not gargle saline as in the previous experiments described hereinabove in Experiments 9, 10, 11, and 12, it appears that saline spraying alone is sufficient for increasing anterior nares testing sensitivity.

[1005] In some applications, any of the techniques described in the report of Experiment 13 above are used in combination with the other techniques described herein.

Experiment 14

[1006] Some of the techniques described hereinabove comprise intranasally dispensing nasal wash fluid into a nasal cavity of a subject; and thereafter, collecting a specimen sample by performing an anterior nares swab, without performing a mid-turbinate swab or a nasopharyngeal swab. Optionally, the method further comprises testing for the presence of a particulate, such as a virus (e.g., SARS-CoV-2), for example using an immunoassay or using isothermal amplification. Experiment 14 provides evidence of the efficacy of these techniques for collecting specimen samples for detection of SARS-CoV-2 using lateral flow immunoassay strips and using isothermal amplification.

[1007] In summary, Experiment 14 found that the sensitivity of lateral flow immunoassay strips improve for anterior nares swabs performed following nasal saline spraying compared to anterior nares swabs performed prior to nasal saline spraying and that, on average, the SARS-CoV-2 antigen band intensity for anterior nares swabs performed following nasal saline spraying were greater than that of anterior flares swabs performed prior to nasal saline spraying, indicating a greater quantity of virus recovered on anterior nares swabs performed following nasal saline spraying than on anterior nares swab performed prior to nasal saline spraying, in support of Experiment 13 hereinabove. Experiment 14 additionally found that the sensitivity of isothermal amplification SARS-CoV-2 detection tests also improve for anterior nares swabs performed following nasal saline spraying compared to anterior nares swabs performed prior to nasal saline spraying.

Detailed Description of Experiment 14: Further Evaluation of Saline Nasal Spraying without Gargling Prior to Anterior Nares Swabbing in SARS-CoV-2 Detection by Immunoassay and Isothermal Amplification and Effects of Head Positioning

Section 1: Materials and Methods

Section 1.1: Patient Population

[1008] This experiment was performed by collecting samples from 14 subjects belonging to the health care service provider Leumit (Tel-Aviv, Israel), who had previously tested positive for SARS-CoV-2. Of the 14 subjects, 6 subjects were subsequently retested the following day (n=20). The clinical study was approved by the investigational review board (IRB) at Assaf Harofeh (Shamir) Medical Center (Tel Aviv, Israel).

Section 1.2: Subject Sampling

[1009] Two alternative sample collection procedures were implemented during subject sampling. First, a unilateral anterior naris swabbing of the right nostril was performed by a healthcare professional. Subjects then sprayed 0.1 mL of normal saline 3-5 times into the right nostril with their heads in a neutral position or slightly tilted back, sniffed in deeply, and then tilted their heads backwards by extending their necks as far as comfortably possible and maintained this head positioning for approximately 15 seconds. The subjects then returned their heads to a neutral position and a second unilateral anterior naris swabbing of the right nostril and then a mid-turbinate swabbing of the right nostril were performed by a healthcare professional. A unilateral anterior naris swabbing of the left nostril was then performed by a healthcare professional. As with the right nostril, subjects then sprayed 0.1 mL of normal saline 3-5 times into the left nostril with their heads in a neutral position or slightly tilted back. Subjects then maintained the neutral or slightly tilted back head positioning for approximately 10 seconds. Lastly, a second unilateral anterior naris swabbing of the left nostril was performed by a healthcare professional.

[1010] Alternatively, following the unilateral anterior naris swabbing of the right nostril and then saline nasal spraying into the right nostril as described above, subjects maintained the neutral or slightly tilted back head positioning for approximately 10 seconds followed by a second unilateral anterior naris swabbing and then a mid-turbinate swabbing of the right nostril by the healthcare professional. Following the unilateral anterior naris swabbing of the left nostril and then saline nasal spraying into the left nostril as described above, subjects then tilted their heads backwards by extending their necks as far as comfortably possible and maintained this head positioning for approximately 15 seconds. The subjects then returned their heads to a neutral position and a second unilateral anterior naris swabbing of the left nostril was performed by a healthcare professional.

[1011] Six subjects were resampled the following day such that the head positioning maintained following saline nasal spraying was reversed for each nostril during resampling compared to during original sampling (Table 11.1).

[1012] Dry swab samples were transported in 15 mL conical tubes (LabCon, USA, Ref #: LC 3136-345-008-9) on ice to Hero Scientific Ltd. (Jerusalem, Israel) for processing. All swabbing was performed with flocked swabs (3BY Ltd., Industrial Park Tefen, Ref #: 3by-001).

Section 1.3: Testing

[1013] Most swab samples were tested by lateral flow immunoassay strips, except for four swab samples: two pairs of unilateral outer naris swab samples that were collected from the left nostril of two subjects with different head positionings were tested by isothermal amplification instead of by lateral flow immunoassay strips (Table 11.1).

[1014] Immunoassays were performed using the BD Veritor™ system for Rapid Detection of SARS-CoV-2 (Becton Dickinson and Company, Maryland, USA, Ref: 256082), in accordance with manufacturer instructions. The BD Veritor™ system for Rapid Detection of SARS-CoV-2 is a chromatographic digital immunoassay intended for the direct and qualitative detection of SARS-CoV-2 nucleocapsid antigens in nasal swabs. In order to determine semi-quantitative differences in viral load between sample types, band intensity was scored on a scale of 0-3, with 0 corresponding to a negative test.

[1015] Isothermal amplification tests were performed using the Lucira™ COVID-19 All-In-One Test Kit system for Rapid Detection of SARS-CoV-2 (Lucira Health, Emeryville, Calif.), in accordance with manufacturer instructions. The Lucira™ COVID-19. All-In-One Test Kit system for Rapid Detection of SARS-CoV-2 is isothermal amplification detection test that utilizes RT-LAMP technology to detect RNA of the N gene for SARS-CoV-2. A positive qualitative result is reported if an RNA signal is detected within 30 minutes and a negative qualitative result is reported if an RNA signal is not detected within 30 minutes. In order to determine semi-qualitative differences in viral load between sample types, the inventors considered the amount of time until results displayed as a relative indicator, with relatively less time until results displayed as an indicator of greater viral load and relatively more time until results displayed as an indicator of lower viral load.

TABLE-US-00014 TABLE 11.1 Overview of Head Positionins Durins Subject Sanwlins, Detection Test Used, and Procedural Deviations Head Detection Test Subject Nostril Position.sup.1 Used  1 Right Neutral Immunoassay Left Back Immunoassay  2 Right Back Immunoassay Left Neutral Immunoassay  3 Right Neutral Immunoassay Left Back Immunoassay  4 Right Back Immunoassay Left Neutral Immunoassay  5 Right Neutral Immunoassay Left Back Immunoassay  6 Right Back Immunoassay Left Neutral Immunoassay  7.sup.§ Right Neutral Immunoassay Left Back Immunoassay  8.sup.§ Right Back Immunoassay Left Neutral Immunoassay  9 Right Neutral Immunoassay Left Back Immunoassay 10 Right Back Immunoassay Left Neutral Immunoassay 11 Right Neutral Immunoassay Left Back Immunoassay 12 Right Back Immunoassay Left Neutral Immunoassay 13.sup.Ω Right Back Immunoassay Left Neutral Immunoassay 14 Right Neutral Immunoassay Left Back Immunoassay  4* Right Neutral Immunoassay Left Back Immunoassay  5* Right Back Immunoassay Left Neutral Isothermal Amplification  6* Right Neutral Immunoassay Left Back Isothermal Amplification  7* Right Back Immunoassay Left Neutral Immunoassay  9* Right Back Immunoassay Left Neutral Immunoassay 11* Right Back Immunoassay Left Neutral Immunoassay iHead positioning immediately following saline nasal spraying. “Back” indicates that the subject maintained a completely tilted back head position for approximately 15 seconds, “Neutral” indicated that the subject maintained a neutral or slightly tilted of back head position for approximately 10 seconds. .sup.ΩSubject did not sniff in after saline nasal spraying. *Resampling subjects with switched head positioning. .sup.§Immunoassay band intensity corresponding to SARS-CoV-2 antigen presence was not scored

Section 2: Results and Discussion

[1016] Immunoassay results were obtained from a total of 38 paired unilateral anterior naris swabs collected before and after saline nasal spraying: 19 paired unilateral anterior naris swabs for each head position (neutral or slightly tilted, and completely tilted back). Isothermal amplification results were obtained from a total of 2 paired unilateral anterior naris swabs collected before and after saline nasal spraying: 1 paired unilateral anterior naris swab for each head position (neutral or slightly tilted, and completely tilted back). Because of expected differences in viral loads between nostrils caused by the nasal cycle, internal comparisons between sampling methodology by pairing nostrils on a patient-by-patient basis were not made, but rather samples from each nostril were treated as samples from independent subjects. Similarly, conclusions were not drawn from comparisons between original samples produced from subjects and samples collected after resampling with alternate methodologies due to temporal changes in nostril dominance, but rather were treated as samples from independent subjects.

Section 2.1: Immunoassay Sensitivity Data

[1017] 11 out of the 19 paired unilateral anterior naris swabs collected from subjects following the sampling protocol in which the head was completely tilted back after saline nasal spraying (hereinbelow, “Head Tilt Protocol”), and 9 out of the 19 paired unilateral anterior naris swabs collected from subjects following the sampling protocol in which the head was neutral or slightly tilted after saline nasal spraying (hereinbelow, “Head Neutral Protocol”), were negative for SARS-CoV-2 antigens on both unilateral anterior naris swabs collected before and after saline nasal spraying. 5 out of the remaining 8 paired unilateral anterior naris swabs collected from subjects following the Head Tilt Protocol, and 4 out of the remaining 10 paired unilateral anterior naris swabs collected from subjects following the Head Neutral Protocol, tested positive for both unilateral anterior naris swabs collected before and after saline nasal spraying.

[1018] Discordant results were reported for 3 paired unilateral anterior naris swabs collected before and after saline nasal spraying from subjects following the Head Tilt Protocol. For all 3 discordant swabs, the unilateral anterior naris swab samples taken after saline nasal spraying were positive for SARS-CoV-2 antigen, and the unilateral anterior naris swab samples taken before saline nasal spraying were negative for SARS-CoV-2 antigen. As seen in Table 11.2, following the Head Tilt Protocol, unilateral anterior naris swabs taken before saline nasal spraying were observed to be 62.5% sensitivity (5/8), while unilateral anterior naris swabs taken after saline nasal spraying had 100% sensitivity (8/8), indicating a 60% increase in detection sensitivity associated with unilateral anterior naris swabbing after saline nasal spraying following the Head Tilt Protocol relative to unilateral anterior naris swabbing performed prior to saline nasal spraying.

[1019] Discordant results were reported for 6 paired unilateral anterior naris swabs collected before and after saline nasal spraying from subjects following the Head Neutral Protocol. For 5 of the 6 discordant results, the unilateral anterior naris swab samples taken after saline nasal spraying was positive for SARS-CoV-2 antigen, and the unilateral anterior naris swab samples taken before saline nasal spraying were negative for SARS-CoV-2 antigen. Conversely, for the single discordant result following the Head Neutral Protocol, the unilateral anterior naris swab sample taken after saline nasal spraying was negative for SARS-CoV-2 antigen, and the unilateral anterior naris swab sample taken prior to saline nasal spraying was positive for SARS-CoV-2 antigen. As seen in Table 11.2, following the Head Neutral Protocol, unilateral anterior naris swabs taken before saline nasal spraying were observed to be 50% sensitivity (5/10), while unilateral anterior naris swabs taken after saline nasal spraying had 90% sensitivity (9/10), indicating an 80% increase in detection sensitivity associated with unilateral anterior naris swabbing after saline nasal spraying following the Head Neutral Protocol relative to unilateral anterior naris swabbing performed prior to saline nasal spraying.

[1020] Because saline nasal spraying increased the sensitivity of the unilateral anterior naris swabs sampled after saline nasal spraying compared to the paired unilateral anterior naris swabs sampled before saline nasal spraying in both Head Neutral and Head Tilt Protocols, it appears that the saline nasal spraying can increase the presence of SARS-CoV-2 virus in the anterior nares independent of head positioning. Furthermore, these observations support previous findings reported in Experiment 13, described hereinabove, that saline nasal spraying benefits SARS-CoV-2 detection on anterior nares swabs in a gargle-independent manner Results reported here appear to support the validity of the Head Neutral Protocol over that of the Head Tilt Protocol. However, given that unilateral anterior naris swabs sampled prior to saline nasal spraying were 25% more sensitive for those sampled during testing of the Head Tilt Protocol relative to those sampled during testing of the Head Neutral Protocol (62.5% and 50%, respectively), despite differences between protocols only effecting samples taken after nasal spraying (unilateral anterior naris swabbing after saline nasal spraying), it is likely that relative differences between protocols are at least partially due to the small sample size. Moreover, and more importantly, as unilateral anterior naris swabbing was performed immediately after returning the head to a normal or slightly tilted back position in the Head Tilt Protocol, it is possible that saline which sampled the virus-rich nasopharyngeal regions had not had sufficient time to drip back into the anterior nares for sampling, as opposed to the Head Neutral Protocol, where the head positioning was maintained for 10 seconds and was thus conducive to saline drippage.

TABLE-US-00015 TABLE 11.2 Immunoassay Sensitivity Head Neutral Protocol Head Tilt Protocol Subject Nostril AN1 AN2 Nostril AN1 AN2 1 Right 0 0 Left 0 0 2 Left 0 0 Right 0 0 3 Right 0 0 Left 0 0 4 Left 0 1 Right 0 0 5 Right 0 0 Left 0 0 6 Left 0 0 Right 0 0 7 Right 0 1 Left 0 1 8 Left 1 0 Right 0 0 9 Right 0 1 Left 1 1 10  Left 0 0 Right 0 0 11  Right 0 0 Left 1 1 12  Left 1 1 Right 0 0 13  Left 1 1 Right 1 1 14  Right 0 1 Left 0 0  4* Right 0 0 Left 1 1  5* Left NA NA Right 0 0  6* Right 0 0 Left NA NA  7* Left 0 1 Right 0 1  9* Left 1 1 Right 1 1 11* Left 1 1 Right 0 1 Sensitivity 50% 90% 62.5% 100% (5/10).sup.i (9/10) (5/8) (8/8) .sup.iSamples were assumed true positive if at least one of the paired unilateral anterior naris swabs before and/or after saline nasal spraying was positive for SARS-CoV-2 antigen. Samples taken from different nostrils from the same subject were assessed as separate subjects. *Resampling subjects with switched head positioning.

Section 2.2: Immunoassay Band Intensity Data

[1021] With regards to the Head Tilt Protocol, for the 5 subjects for whom both of the paired unilateral anterior naris swabs were positive for SARS-CoV-2 antigens, 60% of the time (3/5) band intensity was greater for unilateral anterior naris swabs sampled after saline nasal spraying than unilateral anterior naris swabs sampled before nasal spraying. Similarly, for the 4 subjects for whom both of the paired unilateral anterior naris swabs were positive for SARS-CoV-2 antigens following the Head Normal Protocol, 75% of the time (3/4) band intensity was greater for unilateral anterior naris swabs sampled after saline nasal spraying than unilateral anterior naris swabs sampled before nasal spraying, with equal band intensities for both paired unilateral anterior naris swabs for the remaining subject. The overall trend of stronger band intensities associated with unilateral anterior naris swabs sampled after saline nasal spraying, indicating sampling of more viral material further demonstrates the efficacy of incorporating saline nasal spraying into anterior nares swab sampling methods independent of head positioning.

Section 2.3: Isothermal Amplification Sensitivity Data

[1022] As seen in Table 11.3, one of the two paired unilateral anterior naris swabs collected from subjects following either the Head Tilt Protocol or Head Neutral Protocol tested positive for both unilateral anterior naris swabs collected before and after saline nasal spraying. Discordant results were reported for the one remaining paired unilateral anterior naris swabs collected before and after saline nasal spraying. For this one pair of discordant swabs, the unilateral anterior naris swab sample taken after saline nasal spraying was positive for SARS-CoV-2 antigen, and the unilateral anterior naris swab sample taken before saline nasal spraying was negative for SARS-CoV-2 antigen.

[1023] Furthermore, the one pair of unilateral anterior naris swabs with non-discordant results showed a difference in the amount of time until the isothermal amplification detection test displayed results. The tested unilateral anterior naris swab sample taken before saline nasal spraying displayed results after 19 minutes, while the tested unilateral anterior naris swab sample taken after saline nasal spraying displayed results after only 15 minutes, indicating that the unilateral anterior naris swab sample taken after saline nasal spraying had a relatively greater viral load compared to the unilateral anterior naris swab sample taken before saline nasal spraying.

TABLE-US-00016 TABLE 11.3 Isothermal Amplification Results Time Until Result Subject Nostril Head Position Swab Result Displayed (min) 5* Left Neutral AN1 0 30 (maximum) AN2 1 22 6* Left Back (Tilt) AN1 1 19 AN2 1 15

Section 3: Concluding Remarks

[1024] The evidence presented herein shows that the sampling methods described herein that comprise saline nasal administration (e.g., by spraying) prior to anterior nares swabbing not only increase immunoassay sensitivity, but also antigen band intensities compared to anterior nares swabbing alone, which, for example, may help in examination of immunoassay results even without the aid of an automated immunoassay reader.

[1025] Additionally, the evidence presented herein shows that the sampling methods described herein that comprise saline nasal administration (e.g., by spraying) prior to anterior nares swabbing increase isothermal amplification testing sensitivity and decrease the amount of time until positive results are displayed. As in Experiment 13, subjects did not gargle saline as in Experiments 9, 10, 11, and 12 described hereinabove, indicating that saline spraying alone is sufficient for increasing anterior nares testing sensitivity. This data also suggest the benefit of saline nasal administration prior to anterior nares swabbing independent of head positioning, since sensitivity increased when using a saline nasal spraying protocol involving a neutral head positioning and when using a saline nasal spraying protocol involving a tilted head positioning. However, the evidence presented herein is insufficient for determining which head positioning protocol is superior, since results were likely confounded by procedural variations between examined protocols (i.e., unilateral anterior naris swabbing was performed immediately after returning the head to a normal or slightly tilted back position in the Head Tilt Protocol, so it is possible that saline which sampled the virus-rich nasopharyngeal regions had not had sufficient time to drip back into the anterior nares for sampling, as opposed to the Head Neutral Protocol, where the head positioning was maintained for 10 seconds and was thus conducive to saline drippage).

[1026] In some applications, any of the techniques described in the report of Experiment 14 above are used in combination with the other techniques described herein.

Experiment 15

[1027] Some of the techniques described hereinabove comprise intranasally dispensing nasal wash fluid into a nasal cavity of a subject; and thereafter, collecting a specimen sample by performing an anterior nares swab, without performing a mid-turbinate swab or a nasopharyngeal swab. Optionally, the method further comprises testing for the presence of a particulate, such as a virus (e.g., SARS-CoV-2), for example using quantitative PCR (qPCR). Experiment 15 provides evidence of the efficacy of these techniques for collecting specimen samples for detection of SARS-CoV-2 using qPCR.

[1028] In summary, Experiment 15 found that detection test sensitivity increases when the subject tilts his or her head as far back as comfortably possible after saline nasal administration prior to anterior nares swabbing compared to when saline nasal administration is performed with a head neutral positioning prior to anterior nares swabbing. Experiment 15, unlike Experiment 14, included a wait time between returning the head to a neutral or slightly tilted back position and performing an anterior nares swab sampling, which improved viral load in the sample and improved test sensitivity.

Detailed Report of Experiment 15: Effects of Head Positioning on Saline Nasal Spraying without Gargling Prior to Anterior Nares Swabbing in SARS-CoV-2 Detection by qPCR

Section 1: Materials and Methods

Section 1.1: Patient Population

[1029] This experiment was performed by collecting samples from 4 subjects belonging to the health care service provider Leumit (Tel-Aviv, Israel), who had previously tested positive for SARS-CoV-2. The clinical study was approved by the investigational review board (IRB) at Assaf Harofeh (Shamir) Medical Center (Tel Aviv, Israel).

Section 1.2: Subject Sampling

[1030] A unilateral anterior naris swabbing of the right nostril was performed by a healthcare professional. Subjects then sprayed 0.1 mL of normal saline 3-5 times into the right nostril with their heads in a neutral position or slightly tilted back, sniffed in deeply, and then tilted their heads backwards by extending their necks as far as comfortably possible and maintained this head positioning for approximately 15 seconds. The subjects then returned their heads to a neutral or slightly tilted back position and waited approximately 10 seconds before a second unilateral anterior naris swabbing of the right nostril was performed by the healthcare professional. A mid-turbinate swabbing of the right nostril was then performed by a healthcare professional. A unilateral anterior naris swabbing of the left nostril was then performed by a healthcare professional. As with the right nostril, subjects then sprayed 0.1 mL of normal saline 3-5 times into the left nostril with their heads in a neutral position or slightly tilted back. Subjects then maintained the neutral or slightly tilted back head positioning for approximately 10 seconds. Lastly, a second unilateral anterior naris swabbing of the left nostril was performed by a healthcare professional. A single subject felt unwell and refused the final unilateral anterior nares swabbing (left nostril, after saline nasal spraying).

[1031] All swabs were placed into tubes containing 3 mL of Universal Transport Medium (UTM) (Biological Industries, Israel) and transported on ice to the Leumit Health Services Central Laboratory (Or Yehuda, Israel), for processing via qPCR.

Section 1.3: Molecular Testing

[1032] RNA extraction and qPCR were performed using the Cobas SARS-CoV-2 Test (Roche, Switzerland) on a Cobas 6800 system (Roche, Switzerland), in accordance with manufacturer instructions.

Section 2: Results and Discussion

[1033] Average Ct score difference between unilateral anterior nares swabs collected before saline nasal spraying and unilateral anterior nares swabs collected after saline nasal spraying were greater when subjects tilted their heads back (“Head Tilting Protocol”) after saline nasal spraying compared to when subjects maintained a neutral or slightly tilted back head position (“Head Neutral Protocol”) after saline nasal spraying (Table 12.3).

TABLE-US-00017 TABLE 12.1 aPCR resultsii Ct Scores Subject Sample.sup.i ORF1a/b E 1 Tilt-AN1 36.45 38.08 Tilt-AN2 36.67 40 Neutral-AN1 31.98 33.32 Neutral-AN2 31.46 32.67 2 Tilt-AN1 35.16 36.83 Tilt-AN2 29.66 30.36 Neutral-AN1 29.31 29.75 Neutral-AN2.sup.iii N/A N/A 3 Tilt-AN1 33.0 34.42 Tilt-AN2 31.53 32.38 Neutral-AN1 33.27 34.77 Neutral-AN2 33.6 35.19 4 Tilt-AN1 27.86 28.36 Tilt-AN2 22.85 23.24 Neutral-AN1 27.31 27.8 Neutral-AN2 24.59 24.89 i″Tilt″-swabbing was performed on right nostril, wherein the head was tilted after saline nasal spraying. ″Neutral″-swabbing was performed on the left nostril, wherein the head remained in a neutral or slightly tilted back position following saline nasal spraying. ″AN1″-Unilateral anterior nares swabbing performed prior to saline nasal spraying. ″AN2″-Unilateral anterior nares swabbing performed after saline nasal spraying. .sup.iiNegative PCR results are represented by a Ct score of 40. .sup.iiiSubject refused sampling.

TABLE-US-00018 TABLE 13.2 Ct Score Differences Head Ct Score Difference.sup.i Subject Position.sup.ii ORF1a/b gene E gene 1 Tilt 0.22 1.92 Neutral −0.52 −0.65 2 Tilt −5.5 −6.47 Neutral N/A N/A 3 Tilt −1.47 −2.04 Neutral 0.33 0.42 4 Tilt −5.01 −5.12 Neutral −2.72 −2.91 .sup.iCt score differences were calculated by subtracting Ct scores of unilateral anterior nares swabs performed prior to saline nasal spraying from Ct scores of paired unilateral anterior nares swabbing performed after saline nasal spraying. .sup.ii“Tilt”- swabbing was performed on right nostril, wherein the head was tilted after saline nasal spraying. “Neutral”- swabbing was performed on the left nostril, wherein the head remained in a neutral or slightly tilted back position following saline nasal spraying.

TABLE-US-00019 TABLE 12.3 Average Ct Score Difference Head Average Ct Score Difference Position.sup.i ORF1/a/b gene E gene Tilt −2.94 −2.93 Neutral −0.97 −1.04 .sup.i″Tilt″-swabbing was performed on right nostril, wherein the head was tilted after saline nasal spraying. ″Neutral″-swabbing was performed on the left nostril, wherein the head remained in a neutral or slightly tilted back position following saline nasal spraying.

Section 3: Concluding Remarks

[1034] Although Experiment 14, described hereinabove, may indicate that the Head Tilting Protocol is inferior to the Head Neutral Protocol for virus sampling, it is likely that the addition of a wait period before subjects return their heads to a neutral or slightly tilted position from the tilted back position, and/or the addition of a wait time after the subjects returned their heads to the neutral or slightly tilted back position, before swabbing (which was not included in the sampling protocol for Experiment 14 but was included in the sampling protocol for this Experiment 15) facilitates the dripping of saline from within the nasal cavity into the anterior nares. It therefore appears that a combination of the Head Tilting Protocol and a wait time before subjects return their heads to a neutral or slightly tilted back position, and/or a wait time after the subjects return their heads to the neutral or slightly tilted back position, and performing an anterior nares swab sampling improves viral load in the sample and improves test sensitivity.

[1035] In some applications, any of the techniques described in the report of Experiment 15 above are used in combination with the other techniques described herein.

[1036] In an embodiment, the techniques and apparatus described herein are combined with techniques and apparatus described in one or more of the following patent applications, which are assigned to the assignee of the present application and are incorporated herein by reference: PCT Publication WO 2018/158768 to Fruchter et al.; US Patent Application Publication 2019/0381498 to Fruchter et al.; U.S. Provisional Application 62/727,208, filed Sep. 5, 2018; U.S. Provisional Application 62/727,268, filed Sep. 5, 2018; PCT Publication WO 2020/049566 to Fruchter et al.; PCT Publication WO 2020/049569 to Fruchter et al., and U.S. application Ser. No. 17/270,544 in the national stage thereof; US Patent Application Publication 2021/0102876 to Fruchter et al. U.S. Provisional Application 62/896,295, filed Sep. 5, 2019 U.S. Provisional Application 62/988,145, filed Mar. 11, 2020; U.S. Provisional Application 62/988,259, filed Mar. 11, 2020; U.S. Provisional Application 63/020,723, filed May 6, 2020; U.S. Provisional Application 63/037,707, filed Jun. 11, 2020; U.S. Provisional Application 63/067,535, filed Aug. 19, 2020; U.S. Provisional Application 63/117,294, filed Nov. 23, 2020; U.S. Provisional Application 63/156,843, filed Mar. 4, 2021; U.S. Provisional Application 63/158,005, filed Mar. 8, 2021; U.S. Provisional Application 63/166,378, filed Mar. 26, 2021; U.S. Provisional Application 63/176,565, filed Apr. 19, 2021; PCT Publication WO 2021/044417 to Holtz et al; International Application PCT/IB2021/052055, filed Mar. 11, 2021; and International Application PCT/IB2021/052056, filed Mar. 11, 2012.

[1037] It will be appreciated by persons skilled in the art that the present invention is not limited to what has been particularly shown and described hereinabove. Rather, the scope of the present invention includes both combinations and subcombinations of the various features described hereinabove, as well as variations and modifications thereof that are not in the prior art, which would occur to persons skilled in the art upon reading the foregoing description.