Method for detecting antigens or antibodies to the antigens using EBC and EBC collection device

Abstract

An exhaled breath condensate (EBC) collection device comprising two adjacent parts foldable together and apart from each other, a condensation zone to condense exhaled breath from a subject, the condensation zone being a third part provided between the two adjacent parts and connectable to cooling means arranged in each of the two parts being in cooling relationship with the condensation zone. There is also provided a method for detecting antigens or antibodies to the antigens, the method comprising collecting EBC from a subject using the EBC collection device, performing immunochromatographic testing (ICT) of the collected exhaled breath condensate, and interpreting the immunochromatographic testing results as follows: if the immunochromatographic testing results are positive, the antigens or antibodies to the antigens are detected, or if the immunochromatographic testing results are negative, the antigens or antibodies to the antigens are not detected.

Claims

1. An exhaled breath condensate (EBC) collection device comprising two adjacent parts foldable together and apart from each other, a condensation zone to condense exhaled breath from a subject, the condensation zone being a third part provided between the two adjacent parts and connectable to cooling means arranged in each of the two parts being in cooling relationship with the condensation zone.

2. The device of claim 1, wherein the two adjacent parts and the condensation zone are essentially flat.

3. The device of claim 1, wherein the cooling means are freezer packs or ice packs.

4. The device of claim 1, further comprising an inlet port enabling the subject to breathe into the condensation zone through a detachable mouth-piece.

5. The device of claim 1, wherein the condensation zone comprises an exhaled breath condensate collection area in a bottom area of the condensation zone, enabling the generated breath condensate flow down to the collected area for subsequent collection for performing tests.

6. The device of claim 1, further comprising a perforated label connecting the two adjacent parts in a folded-together state.

7. The device of claim 1, further comprising a breathing indicator that is indicative of a rate of exhaled breath.

8. A method for detecting antigens or antibodies to the antigens, the method comprising: collecting exhaled breath condensate (EBC) from a subject using an exhaled breath condensate collection device comprising two adjacent parts foldable together and apart from each other, a condensation zone to condense exhaled breath from a subject, the condensation zone being a third part provided between the two adjacent parts and connectable to cooling means arranged in each of the two parts being in cooling relationship with the condensation zone, performing immunochromatographic testing (ICT) of the collected exhaled breath condensate, and interpreting the immunochromatographic testing results as follows: if the immunochromatographic testing results are positive, the antigens or antibodies to the antigens are detected, or if the immunochromatographic testing results are negative, the antigens or antibodies to the antigens are not detected.

9. The method of claim 1, wherein the antigens are proteins.

10. The method of claim 1, wherein the method includes using a smartphone-based camera and/or app to perform quantitative assessments.

11. The method of claim 1, wherein the collecting of the EBC from the subject includes respiring by the subject through the EBC collection device at least for 5 minutes.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0075] The present invention is further described in the detailed description which follows, in reference to the drawings wherein:

[0076] FIG. 1 illustrates a chart of different phases of Covid-19 infection.

[0077] FIG. 2 illustrates a general view of the EBC collection device.

[0078] FIG. 3 illustrates widely used blood-based tests.

[0079] FIG. 4 illustrates assembling a mouthpiece to the EBC collection device.

[0080] FIG. 5 illustrates the EBC collection device with the parts in a folded-together state and in foldable apart from each other state.

[0081] FIG. 6 illustrates the EBC collection device in a freezer.

[0082] FIG. 7 illustrates a mouthpiece in a protective plastic bag.

[0083] FIG. 8 and FIG. 9 illustrate the EBC collection device and the mouthpiece.

[0084] FIG. 10 shows using of the EBC collection device by a subject.

[0085] FIG. 11 illustrates removal of the mouthpiece from EBC collection device.

[0086] FIG. 12 illustrates how a perforated label on the right-hand side of the device may be broken.

[0087] FIG. 13 illustrates the EBC collection device with the parts folded apart from each other.

[0088] FIG. 14 illustrates the EBC collection device laid face down with the back part opened.

[0089] FIG. 15 illustrates an enlarged view of a small label covering the fluid collection area of the EBC collection device shown on FIG. 14.

[0090] FIG. 16 illustrates drawing out the required quantity of fluid (e.g. by pipette) for tests.

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0091] The subject-matter of the present application is a method for detecting antigens or antibodies to the antigens, the method comprising collecting exhaled breath condensate (EBC) from a subject using an exhaled breath condensate collection device, performing immunochromatographic testing (ICT) of the collected exhaled breath condensate, interpreting the immunochromatographic testing results as follows: [0092] if positive, the antigens or antibodies to the antigens are detected, or [0093] if negative, the antigens or antibodies to the antigens are not detected.

[0094] In the present invention, the ICT may be applied to all manner of blood ultrafiltrates for non-invasive genetic testing, and/or serologic testing, including in the context of various clinical conditions and disorders.

[0095] ICT testing applied to a blood ultrafiltrate may be aimed at detecting either antigens or antibodies. For example, various cancer antigens, viral antigens, disease related antigens, disease related antibodies, rheumatoid related antibodies, etc. can be detected in this manner. Moreover, immunofluorescent ICT allows for the target antigens and/or antibodies to be quantified. Urusov, et al., (2019) write:

[0096] “Quantitative estimation of the results of immunochromatographic tests can substantially enhance the efficiency of their use in human and veterinary medicine, biosafety, consumer protection, and ecological monitoring. Actually, a row of portable detectors has been developed and implemented with the corresponding software for different kinds of immunochromatographic tests, including assays with optical, magnetic, and electrically conductive labels. While retaining the speed, cost effectiveness, and consumer friendliness of the test systems, these detectors enable detecting and processing results while precluding the subjectivity of their interpretations. New solutions based on the use of general-purpose devices such as office scanners, mobile phones, and smartphones are being developed.”

[0097] In some embodiments, the present invention includes the use of a smartphone-based camera and/or app to achieve quantitative assessments.

[0098] In other embodiments, PCR amplification of viral nucleic acid sequences in the EBC and/or other serological methods detecting antigen or antibody to antigen, as alternative to the ICT, can be also envisaged and covered by the present disclosure.

[0099] Accordingly, the present invention enables rapid and efficient detection of various antigens, including proteins, or antibodies to those antigens, using EBC (or, alternatively, other blood ultrafiltrates such as urine and saliva) and does not require blood sampling.

[0100] Furthermore, an exemplary and novel exhaled breath condensate (EBC) collection device (licensed to be used in the above method) is described, comprising two adjacent parts, wherein the two parts are foldable together and apart from each other, a condensation zone to condense exhaled breath from a subject, the condensation zone being the third part provided between the two parts and connectable to a cooling means arranged in each of the two parts being in cooling relationship with the condensation zone.

[0101] The two foldable parts and the condensation zone part are essentially flat, thereby advantageously making the EBC collection device portable and very compact. The cooling means can be freezer packs or ice packs.

[0102] The EBC collection device further comprises an inlet port to enable a user to breathe into the condensation zone of the device through a detachable, mouth-piece. The condensation zone comprises an exhaled breath condensate collection area in the bottom area of the condensation zone, thereby enabling the generated breath condensate flow down to the collected area for subsequent collection (e.g. by pipette) for performing tests.

[0103] The EBC collection device also comprises a perforated label connecting two parts in a folded-together state.

[0104] The EBC collection device may further comprise a breathing indicator that is indicative of a rate of exhaled breath.

[0105] Operation of the EBC Collection Device:

[0106] The EBC collection device is kept inside the protective plastic bag and placed in a freezer overnight. The mouthpiece is prepared to be in an easily accessible location ready to be used during the tests.

[0107] The EBC collection device is removed from the freezer and is taken out from its protective plastic bag. A mouthpiece is also removed from its protective plastic bag, inserted into the inlet port of the EBC collection device. The inlet port is positioned centrally at the top of the EBC collection device with the breathing indicator to its left.

[0108] The ready-to-use EBC collection device is handed to a subject. The subject respires through the EBC collection device for about 10 minutes, or for 5 minutes at least. If at any point the subject needs to stop to catch his/her breath, the timer should be paused and then recommenced when he/she is ready to do so for the remaining duration of the test. It is important that the subject respires through a nose during inhalation and through a mouth during exhalation. Subject's condition permitting, he/she can be instructed to perform explosive coughs and then take full and deep breaths, filling lungs before expiring with moderate strength.

[0109] In order to access the collected breath condensate, the mouthpiece should first be removed from the EBC collection device for disposal.

[0110] The perforated label on the right hand side of the device should be broken and the parts should be folded apart from each other (appearing as a book pages open) allowing the EBC collection device to stand having the support of said two foldable flat parts. In case, fluid is reluctant to flow into the collection area (if there is an air trap), the collection area can be squeezed to push the air out and to allow the breath condensate to be pulled into the collection area when pressure is removed.

[0111] To access the exhaled breath condensate the EBC collection device is laid face down on a surface and the back part is opened. The small label covering the fluid collection area is peeled off and the required quantity of fluid is drawn out (e.g. by pipette) for tests.

[0112] The immunochromatographic testing is performed using single-use, disposable cartridges or strips which generate detectable coloured end products interpreted as positive or negative.

[0113] Accordingly, in the present invention the EBC instead of blood can be rapidly and efficiently analyzed for the presence of antigens and/or antibodies. Such analysis is less invasive and allows testing at home or in almost any other setting, not only in clinical settings.

[0114] The current methods are useful in any disease or clinical condition in which an ultrafiltrate (e.g., EBC, saliva or urine) can be assessed in conjunction genetic testing. For example, one skilled in the art can apply standard and available algorithms to identify appropriate sequences for PCR testing related to a sequence expressed in cancer or HIV infection.

[0115] The current methods are useful in any disease or clinical condition in which an ultrafiltrate (e.g. EBC, saliva or urine) can be assessed in conjunction serologic testing. For example, one skilled in the art can identify appropriate reporter antibodies for use in ICT, ELISA, or Western blots.