SAMPLE COLLECTING SWAB TOOL AND METHOD FOR DETECTION OF RESPIRATORY PATHOGEN

Abstract

The present invention relates to a sample collecting swab tool and a method for detection of respiratory pathogens. The present invention relates to a swab tool in which two positions for gripping the swab tool, that is, two gripping points, are formed so as to make it easy to precisely control the force on the swab tool according to the sample collection site (e.g., nasal cavity or oral cavity), whereby swab samples can be collected from sample collection sites accurately and easily in an amount sufficient enough to detect respiratory pathogens. In addition, the swab tool of the present invention can be used not only for sampling by experts but also for self-sampling by non-experts, and respiratory pathogens can be detected from collected samples.

Claims

1. A swab device for collecting a specimen, comprising: (a) a head part having a fibrous layer formed to collect a specimen; and (b) a supporting part supporting the head part and having a break point formed on an outer surface; wherein the supporting part comprises a first gripping point and a second gripping point, and the first gripping point is located adjacent to the head part.

2. The swab device according to claim 1, wherein the specimen is a nasal specimen and/or an oral specimen.

3. The swab device according to claim 1, wherein the swab device is a self-sampling device.

4. The swab device according to claim 1, wherein the supporting part is easily cut at the break point.

5. The swab device according to claim 1, wherein the first gripping point and the second gripping point are each formed as an indication part indicating the gripping points.

6. The swab device according to claim 5, wherein the indication part is selected from the group consisting of a dot, a line, a pattern and a grippable shape.

7. The swab device according to claim 6, wherein the grippable shape is a concave shape.

8. The swab device according to claim 1, wherein the first gripping point is a gripping position for collecting a nasal specimen, and the second gripping point is a gripping position for collecting an oral specimen.

9. The swab device according to claim 1, wherein the length from the end of the head part to the first gripping point and the length from the end of the head part to the second gripping point have a ratio of 1:1.2-3.

10. The swab device according to claim 1, wherein the first gripping point is located between the head part and the break point, and the second gripping point is located between the break point and the end of the supporting part.

11. The swab device according to claim 10, wherein the length from the end of the head part to the first gripping point and the length from the end of the head part to the second gripping point have a ratio of 1:1.8-2.5.

12. The swab device according to claim 1, wherein the first gripping point and the second gripping point are located between the break point and the end of the supporting part.

13. The swab device according to claim 12, wherein the length from the end of the head part to the first gripping point and the length from the end of the head part to the second gripping point have a ratio of 1:1.2-1.8.

14. The swab device according to claim 1, wherein the break point of the supporting part is a first break point, the first break point is located between the head part and the first gripping point, the supporting part further comprises a second break point, and the second break point is located between the first gripping point and the second gripping point.

15. The swab device according to claim 14, wherein the length from the end of the head part to the first gripping point and the length from the end of the head part to the second gripping point have a ratio of 1:1.1-1.6.

16.-17. (canceled)

18. The swab device according to claim 14, wherein the length from the end of the head part to the first break point and the length from the end of the head part to the second break point have a ratio of 1:1.8-2.5.

19. The swab device according to claim 1, wherein the overall length of the swab device is 140-180 mm.

20. The swab device according to claim 1, wherein the swab device is a swab device for collecting a nasal specimen and an oral specimen.

21. A kit for collecting a specimen to detect a respiratory pathogen, comprising: (a) the swab device for collecting a specimen of claim 1; and (b) a container comprising a specimen transport medium.

22.-35. (canceled)

36. A method for collecting a specimen to detect a respiratory pathogen, comprising: (a) obtaining a nasal swab specimen and an oral swab specimen by using the kit for collecting a specimen of claim 21; and (b) immersing the nasal swab specimen and the oral swab specimen in a specimen transport medium included in one container.

37.-48. (canceled)

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0200] FIG. 1 shows conventional swab devices for collecting a specimen having different shapes, structures, and/or lengths according to specimen collection sites.

[0201] FIGS. 2, 3 and 4 show swab devices for collecting a specimen according to an embodiment of the present invention.

[0202] FIG. 5 shows a head part of a swab device for collecting a specimen according to an embodiment of the present invention.

[0203] FIGS. 6 to 7 show a shape of a gripping point according to one embodiment of the present invention.

[0204] FIG. 8 illustrates a process of collecting nasal and oral swab specimens using the swab device for collecting a specimen of FIGS. 2 and 3, cutting the swab device in a break point, dipping the swab specimen in a specimen transport medium, and transporting the swab specimen to a testing site.

[0205] FIG. 9 illustrates a process of collecting a nasal and oral swab specimen using the swab device for collecting a specimen of FIG. 4, cutting the swab device in a break point, and dipping the swab specimen in a specimen transport medium to transport the swab specimen to a testing site. The nasal swab specimen collected using the swab device of FIG. 4 is cut in the first break point and then dipped in the specimen transport medium, and the oral swab specimen collected using the swab device of FIG. 4 is cut in the second break point and then dipped in the specimen transport medium.

[0206] FIG. 10 shows an example of collecting a specimen from the nostril.

[0207] FIG. 11 shows an example of collecting a specimen from the oral cavity.

[0208] The present invention will now be described in further detail by examples. It would be obvious to those skilled in the art that these examples are intended to be more concretely illustrative and the scope of the present invention as set forth in the appended claims is not limited to or by the examples.

EXAMPLES

Comparative Example 1: Detection of SARS-CoV-2 by Collecting Nostril Swab Specimen and Oral Swab Specimen

[0209] The nasopharyngeal and oropharyngeal swab specimens were collected by using swab devices for collecting the nasopharyngeal swab specimen and the oropharyngeal swab specimen (Copan Company) included in package 1 of a kit for collecting a specimen among swab devices for collecting a specimen shown in FIG. 1.

[0210] Then, the nasopharyngeal swab specimen and the oropharyngeal swab specimen were immersed together in a clinical virus transport medium (CTM) (Noble Bio) or UTM (Copan) specimen transport medium, which is a balanced salt solution-based buffer. Then, the specimen transport medium was subjected to heat treatment or nucleic acid isolation to obtain a target nucleic acid, which is to be added to RT-PCR reactants.

[0211] To obtain the target nucleic acid by heat treatment, the specimen transport medium in which the nasopharyngeal swab specimen and the oropharyngeal swab specimen were immersed was diluted to three times, and incubated at 98 C. for 3 minutes, thereby obtaining the target nucleic acid (hereinafter, referred to as crude nucleic acid).

[0212] The nucleic acid isolation was carried out using the STARMag 964 Universal Cartridge Kit (Cat. No. 744300.4. UC384, Seegene Inc.) and the automatic nucleic acid extraction system, Microlab NIMBUS (Cat. No. 65415-02, Hamilton). The nucleic acid isolation was conducted according to the manual from the manufacturer of nucleic acid isolation reagents and the operation manual of the system. The nucleic acid isolation was conducted using 300 l of the specimen transport medium in which the nasopharyngeal swab specimen and the oropharyngeal swab specimen were immersed (hereinafter, isolated nucleic acid).

[0213] The RT-PCR reaction using the crude nucleic acid or isolated nucleic acid was performed using Allplex SARS-CoV-2 Assay (Seegene Inc.), which is a multiple one-step RT-PCR product. The Allplex SARS-CoV-2 Assay product is a one-step RT-PCR product for detecting a target gene (E gene) of both Sarbecovirus and SARS-CoV-2 and target genes (RdRP gene, S gene, and N gene) of SARS-CoV-2.

[0214] A reaction mixture of the RT-PCR reaction having a final volume of 20 l was prepared using 5 l of the crude nucleic acid or the isolated nucleic acid, 5 l of enzymes (including chemically modified hot-start Taq polymerase, RTase, UDG, and RI), 5 l of RNase-free Water, and 5 l of SARS2 MOM (Oligo mix).

[0215] Each tube containing the prepared reaction mixture was placed in a real-time thermocycler (CFX96, Bio-Rad), and then reacted at 50 C. for 20 minutes, denatured at 95 C. for 15 minutes, followed by repeating 45 cycles of 10 seconds at 95 C., 15 seconds at 60 C., and 10 seconds at 72 C. The experiment was repeated twice to obtain an average Ct value.

[0216] As a result of the experiment, the use of the isolated nucleic acid obtained from the nasopharyngeal swab specimen and the oropharyngeal swab specimen could obtain a positive signal for SARS-CoV-2. In addition, the use of the crude nucleic acid from the nasopharyngeal swab specimen and the oropharyngeal swab specimen could also obtain a positive signal for SARS-CoV-2, but showed a Ct value delayed by about 1-3 compared with the isolated nucleic acid, indicating a small drop of sensitivity by about 3-6 times.

Example 1: Detection of SARS-CoV-2 by Collecting Nostril Swab Specimen and Oral Swab Specimen

[0217] Using the two swab devices for collecting a specimen shown in FIG. 3 included in package 1 of a kit for collecting a specimen, the nostril swab specimen and the oral swab specimen were collected to detect SARS-CoV-2. The swab device for collecting a specimen of FIG. 3 has a total length of 160 mm, a head part length of 16 mm, and a supporting part length of 144 mm. Further, in the swab device of FIG. 3, lengths from the end of the head part, which is not adjacent to the supporting part, to the break point, to the first gripping point, and to the second gripping point are 60 mm, 80 mm, and 120 mm, respectively.

[0218] The swab device for collecting a specimen of FIG. 3 includes an elliptical (oval) head part having a fiber layer, and the thickness of the head part in a horizontal long axis portion is 5.0 mm and the thickness of the head part in a horizontal short axis portion is 3.0 mm. The fiber layer of the swab device is formed by coating polyester fiber of 3.30-3.35 Dtex with a flocking technique.

[0219] The first gripping point of the swab device was gripped, the swab device was inserted into the nostril by 2 cm, and swabbing was performed while gently rotating for 20 seconds, thereby obtaining a nostril swab specimen. After swabbing in one side of the nostril, swabbing in the other side of the nostril was performed. Then, the second gripping point of the swab device was gripped, and the swab device was applied to the sublingual portion to perform swabbing so that saliva was sufficiently absorbed by the fiber layer of the swab device, thereby obtaining an oral swab specimen.

[0220] Then, in a tube including a specimen transport medium, the swab device obtaining the nostril swab specimen was cut at the first break point, the swab device obtaining the oral swab specimen was cut at the second break point, and the nostril swab specimen and the oral swab specimen were immersed into CTM (Clinical Virus Transport Medium) (Noble Bio Company) or UTM (Copan Company) specimen transport medium, which are balanced salt solution-based buffer, together. Subsequently, a target nucleic acid to be added to the RT-PCR reactant was obtained by heat-treating the specimen transport medium or performing a nucleic acid separation process.

[0221] To obtain the target nucleic acid by heat treatment, the specimen transport medium in which the nostril swab specimen and the oral swab specimen were immersed was diluted to three times, and incubated at 98 C. for 3 minutes, thereby obtaining the target nucleic acid (hereinafter, referred to as crude nucleic acid).

[0222] The nucleic acid isolation was carried out using the STARMag 964 Universal Cartridge Kit (Cat. No. 744300.4. UC384, Seegene Inc.) and the automatic nucleic acid extraction system, Microlab NIMBUS (Cat. No. 65415-02, Hamilton). The nucleic acid isolation was conducted according to the manual from the manufacturer of nucleic acid isolation reagents and the operation manual of the system. The nucleic acid isolation was conducted using 300 l of the specimen transport medium in which the nostril swab specimen and the oral swab specimen were immersed (hereinafter, isolated nucleic acid).

[0223] The RT-PCR reaction using the crude nucleic acid or isolated nucleic acid was performed using Allplex SARS-CoV-2 Assay (Seegene Inc.), which is a multiple one-step RT-PCR product. The Allplex SARS-CoV-2 Assay product is a one-step RT-PCR product for detecting a target gene (E gene) of both Sarbecovirus and SARS-CoV-2 and target genes (RdRP gene, S gene, and N gene) of SARS-CoV-2.

[0224] A reaction mixture of the RT-PCR reaction having a final volume of 20 l was prepared using 5 l of the crude nucleic acid or the isolated nucleic acid, 5 l of enzymes (including chemically modified hot-start Taq polymerase, RTase, UDG, and RI), 5 l of RNase-free Water, and 5 l of SARS2 MOM (Oligo mix).

[0225] Each tube containing the prepared reaction mixture was placed in a real-time thermocycler (CFX96, Bio-Rad), and then reacted at 50 C. for 20 minutes, denatured at 95 C. for 15 minutes, followed by repeating 45 cycles of 10 seconds at 95 C., 15 seconds at 60 C., and 10 seconds at 72 C. The experiment was repeated twice to obtain an average Ct value.

[0226] As a result of the experiment, when the swab device for collecting a specimen of FIG. 3 was used, the positive rate of SARS-CoV-2 was equivalent to that of the swab device for collecting the nasopharyngeal and oropharyngeal specimens used in Comparative Example 1. Meanwhile, the swab device for collecting the nasopharyngeal and oropharyngeal specimens used in Comparative Example 1 took 3-4 times or more time to collect a specimen compared to the swab device for collecting a specimen of FIG. 3. In addition, in the case of using the swab device for collecting a specimen of FIG. 3, self-sampling by a general person who is a non-expert was also possible unlike the swab device used in Comparative Example 1.

[0227] Having described a preferred embodiment of the present invention, it is to be understood that variants and modifications thereof falling within the spirit of the invention may become apparent to those skilled in this art, and the scope of this invention is to be determined by appended claims and their equivalents.