Devices and methods for collecting and analyzing bioaerosol samples

Abstract

A bioaerosol collection device for collecting a bioaerosol sample, comprising: a hollow housing including an inlet, an outlet, and a passage fluidly connecting the inlet and the outlet; and a capture substrate positioned in the passage downstream of the inlet toward the outlet, such that the bioaerosol sample contacts the capture substrate as it flows from the inlet toward the outlet.

Description

Technical Field

[0001] (Cross - Reference to Related Applications) This application claims priority to U.S. Patent Application No. 63 / 136,723, filed on January 13, 2021; U.S. Patent Application No. 63 / 148,195, filed on February 11, 2021; U.S. Patent Application No. 63 / 222,745, filed on July 16, 2021; U.S. Patent Application No. 63 / 224,242, filed on July 21, 2021; U.S. Patent Application No. 63 / 237,909, filed on August 27, 2021; U.S. Patent Application No. 63 / 255,363, filed on October 13, 2021; U.S. Patent Application No. 63 / 283,075, filed on November 25, 2021; and U.S. Patent Application No. 63 / 287,911, filed on December 9, 2021, the disclosures of which are hereby incorporated by reference in their entireties.

Background Art

[0002] This disclosure generally relates to medicine. More particularly, this disclosure relates to devices and methods for collecting and analyzing bioaerosol samples.

[0003] In the medical field, there is a need for devices to collect samples from individuals for testing, analysis, and diagnosis. Sample collection can include invasive methods of sample collection and non - invasive methods of sample collection. After collection, the sample may require further manipulation to prepare the sample for analysis.

[0004] Examples of invasive methods of sample collection include procedures such as surgery and blood sampling. A milder form of invasive methods of sample collection can include swabbing. Invasive methods can cause pain, discomfort, and stress to the patient. Devices used in invasive sample collection methods can also be expensive and may require aseptic handling to prevent sample contamination. Non - invasive methods of collecting samples can be more advantageous than invasive sample collection methods by reducing pain and discomfort in the individual during sample collection.

[0005] Both non-invasive and invasive sample collection may require further handling and processing of the sample for analysis. Sample collection also involves further handling of the sample after collection. For example, a sample collected using a swab requires the transfer of the sample from its original location to a vial, where a portion of the swab containing the collected sample is moved. Cytological samples such as saliva and sputum can be collected in a sample collection vial, and then a portion of the sample is analyzed after all or part of the cytological sample is removed by pipetting. Other liquid samples such as urine and blood are typically collected in a sample collection vial, and then a portion of the sample is analyzed by transferring the sample to another container. The risk of sample contamination arises each time the sample is transferred and can lead to inaccurate results.

[0006] Tissue sample collection may require surgery to collect the sample, depending on the location of the tissue being sampled. Samples such as serum samples require whole blood collection, followed by processing of the whole blood to obtain serum for analytical testing. Alternative sample types are desirable to reduce or eliminate patient discomfort and the complexities of sample collection and handling.

[0007] Therefore, there is a need for alternative non-invasive sample collection devices and methods for collecting samples from individuals. The devices and methods of this disclosure can minimize sample handling and prevent sample contamination. The devices and methods of this disclosure also enable patients to collect samples by evacuating air from the device, without requiring medical professionals. The collected samples can then be transported directly from the patient to a testing facility. Other embodiments of the devices and methods of this disclosure enable patients to perform tests on the collected samples. [Overview of the project]

[0008] This disclosure generally relates to devices and methods for sample collection. In particular, this disclosure relates to devices and methods for collecting air samples from individuals.

[0009] In one embodiment, the present disclosure relates to a bioaerosol collection device for collecting a bioaerosol sample. The bioaerosol collection device includes a hollow housing including an inlet, an outlet, and a passage that fluidly couples the inlet and the outlet, and a capture substrate positioned in the passage downstream of the inlet toward the outlet, wherein the bioaerosol sample comes into contact with the capture substrate as the bioaerosol sample flows from the inlet toward the outlet.

[0010] In one embodiment, the disclosure relates to a bioaerosol collection device for collecting a bioaerosol sample. The bioaerosol collection device comprises a hollow housing including an inlet, an outlet coupled to an analytical device, and a passage fluidly coupling the inlet and the outlet, and a capture substrate disposed in the passage downstream of the inlet, wherein the bioaerosol sample comes into contact with the capture substrate as it flows from the inlet to the outlet and extends through the outlet, with at least a portion of the capture substrate in contact with the sample area of ​​the analytical device.

[0011] In one embodiment, the present disclosure relates to a bioaerosol collection device for collecting a bioaerosol sample. The bioaerosol collection device comprises a capture substrate configured to capture an analyte in a bioaerosol sample; a first bioaerosol permeable protective layer disposed on the upper surface of the capture substrate and covering the upper surface of the capture substrate; and a second bioaerosol permeable protective layer disposed on the lower surface of the capture substrate and covering the lower surface of the capture substrate.

[0012] In one embodiment, the present disclosure relates to a bioaerosol collection device for collecting a bioaerosol sample. The bioaerosol collection device comprises a capture substrate configured to capture an analyte in a bioaerosol sample; a first compartment configured to cover the upper surface of the capture substrate; and a second compartment configured to cover the lower surface of the capture substrate and comprising an analytical device, the analytical device comprising a sample pad that is in fluid contact with the lower surface of the capture substrate. The first and second compartments are configured to be joined together to form a seal. When a buffer solution is introduced onto the capture substrate, the analyte is released from the capture substrate into the buffer solution, and the buffer solution flows to the sample pad of the analytical device.

[0013] In another aspect, the Disclosure relates to a method for collecting and analyzing a bioaerosol sample, comprising: obtaining a bioaerosol sample from a subject using a bioaerosol collection device; obtaining a swab sample from the subject; combining the bioaerosol sample and the swab sample; and analyzing the combined sample. [Brief explanation of the drawing]

[0014] In consideration of the following detailed description, this disclosure will be better understood, and features, aspects, and advantages other than those described above will become apparent. Such a detailed description is referenced to the following drawings. [Figure 1] This figure shows a longitudinal cross-sectional view of an exemplary embodiment of a bioaerosol collection device 1001, which includes a housing 10, a closable inlet 11, a closable outlet 12, a passage 13, and a capture substrate 130. The patient directs an air sample towards the inlet as indicated by arrow 111, and the air flows out as indicated by arrow 112. [Figure 2] This figure shows a longitudinal cross-sectional view of an exemplary embodiment of a bioaerosol collection device, further comprising a band filter and an aperture within the band filter and the capture substrate. [Figure 3] This figure shows a longitudinal cross-sectional view of an exemplary embodiment of a bioaerosol collection device having a closed inlet and outlet configuration using caps. [Figure 4] This figure shows a cross-sectional view of an exemplary embodiment of a bioaerosol collection device having a capture substrate support equipped with optional lateral flow assay (LFA) / vertical flow assay (VFA) attachments and a cap attachment. [Figure 5] The diagram shows a cross-sectional view of an exemplary embodiment of a bioaerosol collection device having a lower cap pressure-operated dropper, and the option of either directly introducing buffer into the bioaerosol collection device or closing the bioaerosol collection device with a cap having a buffer pack that releases buffer from the pack when the cap is positioned or tightened. [Figure 6] This figure shows a longitudinal view (A) and a longitudinal cross-sectional view (B) of an integrated vial tube sample collection and assay device having a tapered inner tube that tapers from a larger inner diameter (D1) proximal to the inlet (proximal to the collection area) to a smaller inner diameter (D2) distal to the inlet. The figure also shows a vent (outlet), an inclined collection substrate with bridging material for directing the sample to the integrated assay component (e.g., LFA / VFA), and a window that allows the user to observe the assay results. [Figure 7] This figure shows a longitudinal view of an exemplary embodiment of a bioaerosol collection device, illustrating the air and reagent flow chambers and capture area at the top of the device. [Figure 8] This figure shows a longitudinal cross-sectional view of an exemplary embodiment of a bioaerosol collection device, illustrating the buffer / reagent components on the surface of the inner wall of the device. [Figure 9]This figure shows a longitudinal cross-sectional view of an integrated bioaerosol collection and assay device and assay process. During the collection process, the target material is vented at the device inlet. Inside, the air sample flows toward and through the capture (collection) substrate where the analyte is collected. Buffer solution is added at the inlet and flows to the capture substrate. After contact with the capture substrate, the collected analyte is transferred via a bridge to the assay component (e.g., LFA / VFA sample pad) to initiate the analysis. A window allows the user to observe the assay results. [Figure 10] This figure shows a cross-sectional view of an exemplary embodiment of an integrated bioaerosol collection and assay device having a conical capture substrate supported by an arbitrary conical capture substrate support and in fluid contact with assay components. The conical configuration concentrates the sample flow into the collection area. [Figure 11] This figure shows a longitudinal cross-sectional view of an exemplary embodiment of a bioaerosol collection device having a conical collection substrate. An assay device (e.g., LFA / VFA) may be an integrated component of the device, or the device may be coupled to the assay device. The airflow moves down the inner tube toward the conical capture substrate and is then directed toward the center point of the capture substrate. The analyte captured by the capture substrate is transferred to the assay device to initiate the analysis. [Figure 12] This figure shows a longitudinal view (A) and a cross-sectional view (B) of an integrated bioaerosol collection device having a conical capture substrate coupled to a lateral flow assay (LFA) / vertical flow assay (VFA). The tubular portion of the device can also be reversibly coupled to the bioaerosol collection device housing. The bioaerosol collection device capture housing can also be reversibly coupled to the LFA / VFA. [Figure 13] This figure shows a longitudinal cross-sectional view of an exemplary embodiment of a bioaerosol collection device having a conical capture substrate and coupled to an LFA / VFA. The analyte can be transferred by introducing buffer using a cap having a buffer pack and / or buffer vial. [Figure 14] FIG. [Figure 14] is a diagram showing an exemplary embodiment of a bioaerosol collection device, and is an enlarged view of the details of the vent (B), the details of the capture substrate (C), and a cross-sectional view of the collection substrate / assay component junction (D). [Figure 15] FIG. is a cross-sectional view showing an exemplary embodiment of an integrated bioaerosol collection device having a "T" - shaped design and an LFA / VFA. [Figure 16A] FIG. [Figure 15] is a photograph showing a 3D - printed two - part bioaerosol collection device having an LFA (FIG. 16A) and a capture housing removed from the tube portion coupled to the LFA. [Figure 16B] FIG. is a photograph showing a 3D - printed two - part bioaerosol collection device having an LFA (FIG. 16A) and a capture housing removed from the tube portion coupled to the LFA. [Figure 17] FIG. [Figure 16A] is a diagram of an exemplary embodiment of a bioaerosol collection device enabling a single - part design that allows for a device having a device that can be used as a vial by covering the top (inlet) and bottom (outlet). The buffer can be introduced from a buffer vial and subsequently capped, and / or the device can be capped using a cap having a buffer pack. [Figure 18] FIG. is a diagram of an exemplary embodiment of a bioaerosol collection device showing a two - part configuration that locks the capture substrate in place and releases the capture substrate by separating the two parts of the device in a coupling mechanism. [Figure 19A] FIG. [Figure 16B] shows different mechanisms for removing the capture substrate by independent movement of the top and bottom, such as twisting (FIG. 19A), sliding (FIG. 19B), and pulling (FIG. 19C). [Figure 19B] FIG. shows different mechanisms for removing the capture substrate by independent movement of the top and bottom, such as twisting (FIG. 19A), sliding (FIG. 19B), and pulling (FIG. 19C). [Figure 19C]A diagram showing different mechanisms for removing a capture substrate by independent movement of the upper and lower parts by torsion (FIG. 19A), sliding (FIG. 19B), pulling (FIG. 19C), etc. [Figure 20A] Diagrams showing different mechanisms for transferring a capture substrate from a bioaerosol collection device, such as using a probe (e.g., a swab or a stick) to push the collection substrate out of the device (FIG. 20A), using a movable internal plunger (FIG. 20B), and using a capture substrate with perforations that allow the capture substrate to be torn when pressure is applied to it (FIG. 20C). [Figure 20B] Diagrams showing different mechanisms for transferring a capture substrate from a bioaerosol collection device, such as using a probe (e.g., a swab or a stick) to push the collection substrate out of the device (FIG. 20A), using a movable internal plunger (FIG. 20B), and using a capture substrate with perforations that allow the capture substrate to be torn when pressure is applied to it (FIG. 20C). [Figure 20C] Diagrams showing different mechanisms for transferring a capture substrate from a bioaerosol collection device, such as using a probe (e.g., a swab or a stick) to push the collection substrate out of the device (FIG. 20A), using a movable internal plunger (FIG. 20B), and using a capture substrate with perforations that allow the capture substrate to be torn when pressure is applied to it (FIG. 20C). [Figure 21A] Diagrams showing the transfer of a capture substrate from a bioaerosol collection device, which can be recovered when dropped from the device (FIG. 21A) and transferred to a vial attached to the device (FIG. 21B). [Figure 21B] Diagrams showing the transfer of a capture substrate from a bioaerosol collection device, which can be recovered when dropped from the device (FIG. 21A) and transferred to a vial attached to the device (FIG. 21B). [Figure 22]This diagram shows the elution of a capture substrate. In A, the capture substrate remains in place, and the patient introduces an air sample (oral / nasal air sample) into the device. In B, a vial is attached to the bottom of the device. The capture substrate can be removed whole or partially for the elution process, allowing the eluate to settle at the bottom. Alternatively, the perforations or adhesives attaching the capture substrate to the device can be mechanically broken by mechanical vibration. Elution buffer can be added, and then the device can be capped. Strong mechanical vibration can be applied to elute the capture substrate and collect the eluted material in the attached vial. In C, the capture substrate remains in the device, and buffer is added. The buffer elutes the contents of the capture substrate and collects them in the attached vial. The vial can then be removed, and the buffer containing the analyte can be transported, stored, and / or analyzed. The vial can also be capped. [Figure 23] This figure shows a combination of a bioaerosol collection device and swab use. In A, the capture substrate remains in place, and the subject blows into the device as instructed. In B, a vial is attached to the bottom of the device. The collection substrate is completely or partially removed using a swab for the elution process, allowing the eluate to settle at the bottom. Elution buffer can be added to the device to simultaneously elute / extract the swab and capture substrate. Alternatively, the perforations or adhesives attaching the capture substrate to the device can be mechanically broken by mechanical vibration. By applying strong mechanical vibration, the swab and capture substrate can be eluted, and the eluted material can be collected in the attached vial. In C, the swab and capture substrate remain inside the device, and elution buffer is added. The elution buffer simultaneously elutes the contents of the swab and capture substrate, and the eluate is collected in the attached vial. The vial can then be removed, and the eluate can be transported, stored, and / or analyzed. The vial can also be capped. [Figure 24]This figure shows an exemplary embodiment of a bioaerosol collection device having a funnel (shown with optional vents) for accommodating vials smaller than the bioaerosol collection device itself. [Figure 25] This figure shows an exemplary embodiment of a bioaerosol collection device, including a cap for introducing a buffer and an integrated assay device (e.g., LFA). [Figure 26] This figure shows an exemplary embodiment of a bioaerosol collection device having a cap for introducing a buffer, an integrated assay device (e.g., LFA), and a swab for analyzing a mixed sample obtained by collecting a bioaerosol sample and a swab sample. [Figure 27] This figure (exploded view) shows a side view and a top view of an exemplary embodiment of a bioaerosol collection device including an upper outer layer, a capture substrate layer, and a lower outer layer. [Figure 28] This figure (exploded view) shows a side view and a top view of an exemplary embodiment of a bioaerosol collection device, which includes an upper outer layer, a capture substrate layer, and a lower outer layer, and further includes a protective layer (filter) for protecting the upper and lower surfaces of the capture substrate. [Figure 29] This figure shows a top view of an exemplary embodiment of a bioaerosol collection device, showing the capture region and folds in the outer layer that help remove the outer layer to expose the capture substrate and then process it for analysis. [Figure 30] This is a translucent diagram illustrating an exemplary embodiment of a bioaerosol collection device, showing a capture substrate holder that supports the capture substrate against the air force applied when the user directs a bioaerosol sample into the collection area, and that allows the user to handle the capture substrate without direct contact with it. [Figure 31] This is a translucent diagram illustrating an exemplary embodiment of a bioaerosol collection device, showing an outer layer that covers only a portion of the capture substrate layer and the capture substrate holder. The device edges that come into contact with the user's skin may be flat or arc-shaped. [Figure 32]This is an exploded view (side view and top view) of an exemplary embodiment of a bioaerosol collection device including an airflow ring / tube for directing a bioaerosol sample to a capture substrate region, the capture substrate layer including a capture substrate holder and a capture substrate. [Figure 33] This is an exploded view (side view and top view) of an exemplary embodiment of a bioaerosol collection device. The capture substrate layer includes a capture substrate and a capture substrate holder. The capture substrate holder is releasably connected to a support and is configured to be detached from the support in order to remove the capture substrate from the rest of the bioaerosol collection device. [Figure 34] This is an exploded view (side and top view) of an exemplary embodiment of a bioaerosol collection device. The capture substrate layer includes a teardrop-shaped capture substrate and a capture substrate holder. The capture substrate holder is releasably connected to a support and configured to be detached from the support in order to remove the capture substrate from the rest of the bioaerosol collection device. [Figure 35] This is an exploded view (side and top view) of an exemplary embodiment of a bioaerosol collection device. The capture substrate layer includes a teardrop-shaped capture substrate and a capture substrate holder. The capture substrate holder is releasably connected to a support and configured to be detached from the support to remove the capture substrate from the rest of the bioaerosol collection device. The exemplary embodiment also includes upper and lower tabs located on the upper and lower surfaces of the capture substrate layer. A protective layer protects the capture substrate from contamination and physical damage and can filter the bioaerosol sample. [Figure 36] These are exploded views (side and top views) of an exemplary embodiment of a bioaerosol collection device. The outer layer may include folds that facilitate the removal (separation) of the outer layer to expose the inner layer and allow the captured substrate to be handled for transport and / or analysis. [Figure 37] This is an exploded view (side and top view) of an exemplary embodiment of a bioaerosol collection device, showing the arc-shaped edges of the device intended to conform to the user's skin surface. [Figure 38]This figure shows an exemplary embodiment of the capture layer of a bioaerosol collection device. [Figure 39] This figure shows an exemplary embodiment of a trapping substrate holder, along with an exemplary embodiment of a trapping substrate holder configured to be releasably connected to a support and disconnected from the support. [Figure 40] This figure shows an exemplary embodiment of a bioaerosol collection device, comprising a "clamshell" component including an upper part having a buffer pack and a lower component containing an analytical device (LFA / VFA). [Figure 41] This figure shows a translucent image of a top view and a bottom view of a clamshell embodiment, with the capture substrate inside a closed clamshell for analysis. [Figure 42] The figures show a top view and a bottom view of a clamshell embodiment, in which the capture substrate is located inside a closed clamshell for analysis, and the capture substrate is shown to be coupled to a support layer and to include an upper tab and a lower tab. [Figure 43] This figure shows a top view and a bottom view of a clamshell embodiment, where the capture substrate is inside a closed clamshell and the support has been removed from the holder for analysis. [Figure 44] This is a photographic image of a bioaerosol collection device. [Figure 45] This is a photographic image of a bioaerosol collection device, showing the holder connected to a support and the holder removed from the support (with the capture substrate attached). [Figure 46] This figure shows an exemplary embodiment of a punch-design bioaerosol collection device. The application of force to the upper or lower outer layer moves the capture substrate, allowing the capture substrate to be transported and processed for analysis. [Figure 47] This figure shows an exemplary embodiment of a bioaerosol collection device with a punch design configured to cover the capture area. [Figure 48]This figure shows an exemplary embodiment of a bioaerosol collection device including an airflow adapter. The airflow adapter is configured to receive the bioaerosol collection device, which advantageously allows different users to direct bioaerosol samples to the same bioaerosol collection device by using different airflow adapters. [Figure 49] This figure shows an exemplary embodiment of a bioaerosol collection device, including an airflow adapter configured to allow collection of bioaerosol samples from both nostrils of the user. [Figure 50] This figure illustrates an exemplary embodiment of a bioaerosol collection device, which includes a fluid-coupled buffer pack on a capture substrate that is fluid-coupled to a sample pad of an analytical device. [Figure 51] This figure shows an exemplary embodiment of a bioaerosol collection device in which a capture substrate is coupled with an analytical device. After bioaerosol sample collection, the capture substrate is placed in a vial containing a reagent buffer to initiate analysis via the analytical device. [Figure 52A] This figure shows the collection of a bioaerosol sample (Figure 52A) and the subsequent elution of a swab sample used to elute the bioaerosol sample (Figure 52B). The combined swab and bioaerosol sample are analyzed by an analytical device coupled to the bioaerosol collection device (Figure 52C). [Figure 52B] This figure shows the collection of a bioaerosol sample (Figure 52A) and the subsequent elution of a swab sample used to elute the bioaerosol sample (Figure 52B). The combined swab and bioaerosol sample are analyzed by an analytical device coupled to the bioaerosol collection device (Figure 52C). [Figure 52C] This figure shows the collection of a bioaerosol sample (Figure 52A) and the subsequent elution of a swab sample used to elute the bioaerosol sample (Figure 52B). The combined swab and bioaerosol sample are analyzed by an analytical device coupled to the bioaerosol collection device (Figure 52C). [Figure 53A]This figure shows the process used to combine a bioaerosol sample and a swab sample in an integrated bioaerosol collection device. Steps 1 and 2 involve obtaining a bioaerosol sample (oral and / or nasal cavity) and a swab sample from the subject (Figure 53A). In step 2, the swab is introduced into the bioaerosol collection device, and a buffer solution is introduced to elute the analyte from the swab and the capture substrate. The analyte is then transferred to the assay device. [Figure 53B] This figure shows the process used to combine a bioaerosol sample and a swab sample in an integrated bioaerosol collection device. Steps 1 and 2 involve obtaining a bioaerosol sample (oral and / or nasal cavity) and a swab sample from the subject (Figure 53A). In step 2, the swab is introduced into the bioaerosol collection device, and a buffer solution is introduced to elute the analyte from the swab and the capture substrate. The analyte is then transferred to the assay device. [Figure 54A] This figure shows the process of combining a swab and a bioaerosol sample. Steps 1 and 2 involve obtaining a bioaerosol sample (oral and / or nasal cavity) and a swab sample from the subject (Figure 54A). Step 3 involves adding a buffer solution to a dropper vial (Figure 54B). Step 4 involves using a swab to transfer the capture substrate from the bioaerosol collection device to the dropper vial (Figure 54C). Step 5 involves removing the swab and using the dropper vial to transfer the droplet of the combined sample to an analytical device for analyte detection. [Figure 54B] This figure shows the process of combining a swab and a bioaerosol sample. Steps 1 and 2 involve obtaining a bioaerosol sample (oral and / or nasal cavity) and a swab sample from the subject (Figure 54A). Step 3 involves adding a buffer solution to a dropper vial (Figure 54B). Step 4 involves using a swab to transfer the capture substrate from the bioaerosol collection device to the dropper vial (Figure 54C). Step 5 involves removing the swab and using the dropper vial to transfer the droplet of the combined sample to an analytical device for analyte detection. [Figure 54C]This figure shows the process of combining a swab and a bioaerosol sample. Steps 1 and 2 involve obtaining a bioaerosol sample (oral and / or nasal cavity) and a swab sample from the subject (Figure 54A). Step 3 involves adding a buffer solution to a dropper vial (Figure 54B). Step 4 involves using a swab to transfer the capture substrate from the bioaerosol collection device to the dropper vial (Figure 54C). Step 5 involves removing the swab and using the dropper vial to transfer the droplet of the combined sample to an analytical device for analyte detection. [Figure 54D] This figure shows the process of combining a swab and a bioaerosol sample. Steps 1 and 2 involve obtaining a bioaerosol sample (oral and / or nasal cavity) and a swab sample from the subject (Figure 54A). Step 3 involves adding a buffer solution to a dropper vial (Figure 54B). Step 4 involves using a swab to transfer the capture substrate from the bioaerosol collection device to the dropper vial (Figure 54C). Step 5 involves removing the swab and using the dropper vial to transfer the droplet of the combined sample to an analytical device for analyte detection. [Figure 55] This figure shows the combination of swab sample and bioaerosol sample, obtained by separately eluting the swab and the captured substrate, and then combining the swab sample with the bioaerosol sample to form a composite eluate. [Figure 56] This figure shows the combinations of swab samples and bioaerosol samples obtained by eluting the bioaerosol capture substrate and swab in the same vial. [Figure 57] This figure shows the combination of swab sample and bioaerosol sample, obtained by first eluting the swab and then eluting the bioaerosol-capturing substrate using the swab eluent. [Figure 58] This graph shows that combining samples leads to better detection of SARS-CoV-2 (adapted from Jarvis and Kelley, Scientific Reports, 11, 9221 (April 28, 2021)). [Figure 59]This table summarizes the results of the analysis of nasal air samples collected using an exemplary device embodiment. [Modes for carrying out the invention]

[0015] Unless otherwise specified, all scientific and technical terms used herein have the same meanings as those generally understood by those skilled in the art in which this disclosure pertains. Any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this disclosure, but preferred methods and materials are described below.

[0016] A device and method for collecting bioaerosol samples from a subject are disclosed. In some embodiments, the device is coupled with an analytical device that advantageously enables the collection and analysis of bioaerosol samples alone or in combination with other sample types (e.g., swabs, sputum, lavage solutions, aspirates, etc.). Samples can be collected from a subject by a medical professional, or by the subject with or without the assistance of a medical professional.

[0017] As used herein, “bioaerosol sample” is used in accordance with its ordinary meaning as understood by those skilled in the art, and means an airborne collection of biological material. Bioaerosol samples may include cells, cell fragments, fungal spores, fungal hyphae, viruses, proteins, nucleic acids, other biological substances, and chemical substances.

[0018] As used herein, “capture substrate” (and / or “collection substrate”) refers to a surface or material that collects material contained in a bioaerosol sample. The capture substrate is designed to capture (collect and / or capture) the analyte of interest. The pore size of the capture substrate is selected based on the size of the analyte. Generally, the pore size of the capture substrate is smaller than the size of the analyte of interest so that the analyte does not pass through the capture substrate. The capture substrate may also include a range of pore sizes. The capture substrate favorably concentrates the collected sample so that the amount of reagent required to perform the analysis of the sample is minimized. The concentration and collection function of the capture substrate also improves test sensitivity. The capture substrate may also be designed to release the analyte of interest. For example, the capture substrate may be designed so that the analyte collected by the capture substrate can be eluted from the capture substrate using a buffer that washes the analyte away from the capture substrate, or a buffer that dissolves the capture substrate, thereby releasing the analyte. The capture substrate may also be designed to allow the analyte of interest to move within the capture substrate (e.g., from one area of ​​the capture substrate to another). The capture substrate can also be configured to allow the transfer of the target analyte from the collection substrate to the test substrate. For example, the capture substrate may include pores, channels, grooves, processing areas, fiber types, and mixtures of materials, through which the analyte moves and / or along which the analyte and / or the carrier containing the analyte move away from the capture substrate. For example, the analyte can move through channels (e.g., microchannels) in the capture substrate to the test substrate or another vial.

[0019] The capture substrate can have different shapes. Suitable shapes include circular, disk, elliptical, conical, egg-shaped, teardrop-shaped, square, rectangular, triangular, and basket-shaped. Generally, the shape of the capture substrate is similar to the shape of the airflow channel of the device. The shape of the capture substrate may be similar to the shape of the device designed to hold the capture substrate. The capture substrate can be oriented within the device. For example, as shown in Figures 1-5, the capture substrate is oriented perpendicular to the airflow. As shown in Figures 6-9, the capture substrate is oriented at an angle to the airflow. As shown in Figures 10-15, the capture substrate is oriented perpendicular to the airflow, but the shape of the capture substrate is conical, with a portion of the capture substrate oriented at an angle to the airflow. Conical capture substrates are particularly suitable for focusing sample material toward the center point of the substrate. The capture substrate can be molded to the shape of the device designed to hold the capture substrate, similar to how a coffee filter fits into a coffee filter holder. The capture substrate may further include folds, creases, and other structures to increase the surface area of ​​the capture substrate.

[0020] A capture substrate without an outer layer structure is flexible and can be folded to conform to various shapes and easily positioned inside a vial. The capture substrate is appropriately sized to cover the entire pores (air channels) of the upper and lower outer layers and to reduce or prevent patient exhalation from passing through the capture substrate. The size of the capture substrate is preferably the minimum necessary to cover the air channels and hold the capture substrate in place.

[0021] The capture substrate may also include perforations. Perforations may be included to remove the capture substrate from other layers. Perforations may also be included to remove a portion of the capture substrate, thus allowing only a portion of the capture substrate to be tested further, for example, allowing the remaining portion of the capture substrate to be stored for later testing. Testing a smaller portion of the capture substrate rather than the entire substrate can increase the sensitivity of the test.

[0022] The capture substrate may include "slits" to increase its adaptability so as to easily fit a range of vial sizes, and for comfort and effectiveness in swab the nostrils. The slits may be radial from the center of the substrate or parallel to the capture substrate holder.

[0023] The capture substrate may further include an aperture that allows a user to insert a sample handling device, such as a pipette tip, through the capture substrate. The aperture may also function to reduce the pressure of the airflow over the capture substrate so that it does not detach when an individual directs air into the inlet. While the pressure on the capture substrate can be released and / or reduced by the aperture, the capture substrate can be fabricated to have sufficient strength to withstand the pressure generated when an individual directs air into the inlet of a device. Furthermore, the capture substrate may be deformed by air pressure. The materials used to fabricate the capture substrate, and the method used to secure the capture substrate in place within the housing, will prevent the capture substrate from moving while allowing sufficient airflow through it.

[0024] In certain embodiments, the capture substrate is configured for the further purpose of being used as a swab. By using a flexible bioaerosol capture substrate material to swab the nose (and / or mouth) and combine it with a bioaerosol and a transnasal (and / or oral) sample, the concentration of the analyte can be increased. For example, all or part of the edge of the capture substrate can contain swab material to increase the surface area for swabing. The capture substrate may include a first layer of a first material for collecting analytes in a bioaerosol sample and a second layer containing swab material.

[0025] The capture substrate can be bonded (or attached) to the capture substrate handle (loop and / or frame), filter layer(s), and outer layer using adhesive. Adhesive around the top and bottom of the air channel may be useful to ensure that air does not escape laterally and passes only through the capture substrate. A suitable adhesive is inert and does not interfere with molecular or antigen testing.

[0026] The capture substrate is preferably made from synthetic fibers, natural fibers, and combinations thereof. Fibers used to form the capture substrate include hydrophobic fibers, hydrophilic fibers, and combinations thereof. Examples of hydrophobic fibers include polylactone, poly(caprolactone), poly(L-lactic acid), poly(glycolic acid), similar copolymers poly(alkyl acrylate), polybutadiene, polyethylene, polystyrene, polyacrylonitrile, polyethylene(terephthalate), polysulfone, polycarbonate, poly(vinyl chloride), and combinations thereof. Examples of hydrophilic fibers include linear poly(ethyleneimine), cellulose, cellulose acetate and other grafted cellulose derivatives, poly(hydroxyethyl methacrylate), poly(ethylene oxide), polyvinylpyrrolidone, poly(acrylic acid), poly(ethylene glycol), poly(vinyl alcohol), poly(vinyl acetate), poly(acrylamide), protein, poly(vinylpyrrolidone), poly(styrene sulfonate), and combinations thereof.Other suitable fiber materials include, for example, acrylonitrile / butadiene copolymer, cellulose, cellulose acetate, chitosan, collagen, DNA, fibrinogen, fibronectin, nylon, poly(acrylic acid), poly(chlorostyrene), poly(dimethylsiloxane), poly(etherimide), poly(ethersulfone), poly(ethyl acrylate), poly(ethyl vinyl acetate), poly(ethyl-co-vinyl acetate), poly(ethylene oxide), poly(ethylene terephthalate), poly(lactic acid-co-glycolic acid), poly(methacrylic acid) salt, poly(methyl methacrylate), poly(methylstyrene), poly(styrene sulfonic acid) salt, poly(styrene sulfonyl fluoride), poly(styrene-co-acrylic acid) salt Examples include lilonitrile, poly(styrene-co-butadiene), poly(styrene-codivinylbenzene), poly(vinyl acetate), poly(vinyl alcohol), poly(vinyl chloride), poly(vinylidene fluoride), polyacrylamide, polyacrylonitrile, polyamic acid (PAA), polyamide, polyaniline, polybenzimidazole, polycaprolactone, polycarbonate, polydimethylsiloxane-co-polyethylene oxide, polyetheretherketone, polyethylene, polyethyleneimine, polyimide, polyisoprene, polylactide, polypropylene, polystyrene, polysulfone, polyurethane, polyvinylpyrrolidone, protein, SEBS copolymer, silk, and styrene / isoprene copolymer.Polymer blends include, for example, poly(vinylidene fluoride)-blend-poly(methyl methacrylate), polystyrene-blend-poly(vinyl methyl ether), poly(methyl methacrylate)-blend-poly(ethylene oxide), poly(hydroxypropyl methacrylate)-blend-poly(vinylpyrrolidone), poly(hydroxybutyrate)-blend-poly(ethylene oxide), protein blend-polyethylene oxide, polylactide-blend-polyvinylpyrrolidone, polystyrene-blend-polyester, polyester-blend-poly(hydroxyethyl methacrylate), poly(ethylene oxide)-blend-poly(methyl methacrylate), and poly(hydroxystyrene)-blend-poly(ethylene oxide). The fibrous material used to form the capture substrate may be selected to transfer the analyte (including the carrier containing the analyte) from one part of the capture substrate to another part of the capture substrate. The fibrous material used to form the capture substrate may be selected to transfer the analyte (including the carrier containing the analyte) from one part of the capture substrate to the outside of the capture substrate and / or to the analyte substrate and / or collection vial.

[0027] Another suitable trapping substrate may be an electret (including thermoelectrets and fibrillated electret films). An electret is a dielectric material having a quasi-permanent charge or dipole polarization. Electrets can be obtained from commercially available sources. Electrets can be prepared by heating a material and simultaneously exposing it to an electric field, thereby orienting many of the dipoles in the material in a favorable direction. After heating, the material is "frozen" and its electric dipole positions can be maintained for a long period of time. Suitable materials for preparing electrets include, for example, organic materials such as ebonite, naphthalene, polymethyl methacrylate, and many polymers, as well as inorganic materials such as sulfur, quartz, glass, steatite, and some ceramics, and materials that can currently be used to produce thermoelectrets. Electret fiber films are particularly preferred. Suitable polymer electrets also include poly(L-lactic acid) electrets and polypropylene. Polyvinylidene fluoride (PVDF) / polytetrafluoroethylene (PTFE) NP electret nanofiber films can be formed by electrospinning. Fibrillated electret films, such as those by van Turnhout (U.S. Patent No. 3,998,916), are also suitable. Suitable materials for use as capture substrates include the biosampling materials disclosed by Kanzer. Angadjivand's U.S. Patent No. 6,119,691 discloses electret filter materials. Charged electrets interact with the analyte of interest to capture it. The application of extraction / elution buffer causes a short circuit of charge, releasing all or substantially all of the analyte from the electret capture substrate material.

[0028] Suitable capture substrate materials include those that are soluble and / or liquid-soluble. For example, cellulose acetate nanofibers that dissolve upon contact with a liquid. It should be understood that the entire capture substrate and / or a portion thereof may be soluble or liquid. Advantageously, the capture substrate itself can dissolve, completely liberating all sample material into the eluent, eliminating the need for a removal process to remove the analyte from the capture substrate or any substrate material. Preferably, the capture substrate can be fabricated to react with the sample (e.g., particles, cells, DNA, RNA, etc.) upon dissolution to stabilize it or perform some other service detection. Preferably, the capture substrate may be inert before dissolution.

[0029] In certain embodiments, the capture substrate is processed to transport, elute, and / or extract and / or remove the analyte of interest from the capture substrate. In other embodiments, the analysis does not require the removal or extraction of the analyte from the capture substrate. For example, the capture substrate can be analyzed by adding a reaction solution (e.g., buffer and / or water) to the device that produces a colorimetric reaction indicating the presence or absence of the analyte. In another embodiment, the capture substrate is placed in a reaction solution (e.g., buffer and / or water) by which the capture substrate dissolves. After the capture substrate has dissolved, the analyte to be detected is released into the reaction solution, which can be tested directly by adding other reagents to the device and / or transferring all or part of the solution to another reaction medium such as a vial, tube, membrane, or slide.

[0030] The capture substrate can also be protected on all sides by additional layers such as filters and protective layers. The capture substrate can also be left free from protective layers. The protective layer and the capture substrate can be made from different materials designed for specific purposes (e.g., protection and sample collection). The capture substrate can be coated with reagents to hold the analyte load collection. The inner capture substrate and / or protective layer(s) may also be coated with reagents to stabilize the analyte. The protective layer may be an air-permeable contact protective coating.

[0031] As used herein, “buffer solution” refers to the components used to stabilize, transport, extract, elute, and / or detect an analyte. For example, an analytical reagent may include buffer components, salts, dNTPs, oligonucleotide primers, polymerases, reverse transcriptases, and combinations thereof. Analytical reagents can be appropriately lyophilized in liquid form, gel form, and combinations thereof. The reagent layer containing the analytical reagent may be separated by other layers of the collection substrate by a coating to prevent the test reagent from coming into contact with and / or being activated on the capture substrate until the capture substrate is processed for analysis. The analytical reagent can be activated, for example, by placing the capture substrate (and / or the collection layer having a soluble layer containing the test reagent) in a liquid medium such as a buffer solution containing water, thereby causing the coating to dissolve and release the analytical reagent, which can then also dissolve in the buffer solution. The coating may be meltable, thereby allowing the temperature to be adjusted so that the coating melts and releases the analytical reagent, forming a mixture in which the analyte can be detected. The reagent may also be in the form of reagent-containing microparticles and / or beads. Suitable reagents include salts, pH buffers, transport media, preservatives, capture reagents, analytical reagents, detection reagents, eluents, antibacterial agents such as silver-containing antimicrobial agents and antimicrobial polypeptides, analgesics such as lidocaine, antibiotics such as neomycin, thrombus-forming compounds, nitric oxide-releasing compounds such as cydononimine and NO complexes, bactericidal compounds, fungicidal compounds, bacteriostatic compounds, analgesic compounds, other pharmaceutical compounds, adhesives, fragrances, odor-absorbing compounds, preservatives, RNAse inhibitors, protease inhibitors, and nucleic acids including deoxyribonucleic acid, ribonucleic acid, and nucleotide analogs. Other suitable reagents include capture reagents, such as antibodies that specifically bind to the analyte of interest, and ligands that specifically bind to the analyte of interest (e.g., surface molecules such as sugars and glycoproteins). Capture reagents can be covalently or noncovalently bonded to the collection substrate by a linker.For example, any suitable linker can be used, such as organic molecules such as polymers or copolymers (e.g., substituted or unsubstituted polyalkylene glycols such as polyethylene glycol) and / or biomolecules such as bovine serum albumin. During preparation, the reagent can be introduced into a housing in a liquid medium, and then the liquid portion of the medium can be evaporated to dry or freeze-dry the reagent. Afterwards, the user can rehydrate the dried reagent by adding a liquid medium, and once rehydrated, the reagent is available for its intended purpose. As described herein, analytical reagents can be appropriately freeze-dried in liquid form, gel, and combinations thereof. Analytical reagents can be activated as described herein by adding a liquid medium such as a buffer containing water, thereby dissolving the coating and releasing the analytical reagent, which can then also dissolve in the buffer. The coating may be fusible, so that the temperature can be adjusted so that the coating melts and releases the reagent, forming a mixture in which the analyte can be detected. The reagent coating may contain reagent-containing microparticles and / or beads. Suitable transport media include, for example, viral transport medium (VTM), amies transport medium, and sterile saline. Transport media may contain, for example, RNAse inhibitors, DNase inhibitors, protease inhibitors, preservatives, stabilizers, and other reagents for sample preservation. Once placed in a transport container, the collected and combined samples can be stored for subsequent analysis or processed for analysis. Processing may include dissolution, extraction, and other known processing steps for analyzing the mixed sample for analytes.

[0032] As disclosed herein, adhesives can be used to “bond” the capture substrate to a position within the device (e.g., to a shelf and / or capture substrate holder that contacts the edge of the capture substrate). Additionally, or alternatively, the capture substrate can be “locked” to a position within the device where two parts of the device are reversibly joined to hold the capture substrate (e.g., a “clamshell” arrangement). Additionally, or alternatively, the capture substrate can be bonded to a capture substrate holder using fusion bonding. Additionally, or alternatively, the capture substrate can be bonded to a capture substrate holder using fasteners such as rivets, pins, or screws.

[0033] When in use, the bioaerosol collection device is designed to capture or collect analytes contained in a bioaerosol sample. Suitable analytes may be viruses, bacteria, nucleic acids (e.g., DNA and / or RNA), proteins, chemicals, and combinations thereof. As described herein, the pore size of the capture substrate is designed so that the analyte of interest is captured by the capture substrate. If the capture substrate includes an aperture that allows the user to insert the device into the device, it should be understood that a certain amount of analyte may flow through the analyte and therefore not be captured by the capture substrate. However, a capture substrate with an aperture will capture the analyte of interest from the air flowing through the capture substrate.

[0034] The analytes include microorganisms, biomolecules, and chemical molecules. Of particular interest are pathogens. As used herein, “pathogen” refers to microorganisms such as bacteria, fungi, and viruses. The term “pathogen” also refers to viroids, prions, and proteins.

[0035] Suitable analytes are contained in airborne gases and aerosol droplets released by the user. Suitable analytes include microorganisms, chemicals, proteins, nucleic acids, and combinations thereof. Suitable microorganisms include bacteria and viruses.

[0036] Particularly relevant microorganisms include pathogens. The term "pathogen" is used, in its usual sense, to refer to bacteria, viruses, and other microorganisms that directly or indirectly cause disease. Exemplary pathogens include, for example, Yersinia, Klebsiella, Providencia, Erwinia, Enterobacter, Salmonella, Serratia, Aerobacter, Escherichia, Pseudomonas, Shigella, Vibrio, Aeromonas, Streptococcus, Staphylococcus, Micrococcus, Moraxella, Bacillus, Clostridium, Corynebacterium, Salmonella typhi, Francisella, Haemophilus, Bacteroides, Listeria, Erysipelothrix, Acinetobacter, Brucella, Pasteurella, Flavobacterium, and Fusococcus. Bacterium, Streptobacillus, Callimatobacterium, Legionella, Treponema, Borrelia, Leptospira, Actinomyces, Nocardia, Rickettsia, Micrococcus, Mycobacterium, Neisseria, Campylobacter, pathogenic viruses, such as papillomavirus, parvovirus, adenovirus, herpesvirus, vaccine virus, arenavirus, coronavirus (SARS-CoV-2, coronavirus 229E, coronavirus HKU1, coronavirus NL63, etc.) Navirus (OCL43), rhinovirus, polynuclear respiratory virus, influenza virus, picornavirus, paramyxovirus, reovirus, retrovirus, rhabdovirus, human immunodeficiency virus (HIV), tanya, hymenorepsis, diphyllobacterium, echinococcosis, hypertrophic fluke infection, dysplasia, fluke infection, hepatofluidosis, phyllohelidosis, lung fluke, schistosomiasis, pinworm infection, whipworms, roundworms, hookworms, Necatol, Bukeleria, burgi, loa, onchocerciasis, dracuncurus, naegleria, acanthus Soameba, malaria parasitic infection, trypanosomiasis, leishmaniasis, toxoplasmosis, dysentery amoeba, giardiasis, isosporiasis, cryptosporidiosis, microsporidiasis, strongyloidiasis, trichinellosis, for example, tinea, histoplasmosis, blastomycosis, aspergillosis, cryptococcosis, sporotrichosis, coccidioidomycosis, paracoccidioidomycosis, mucormycosis, candidiasis, tinea, protothecasis, pityriasis, mycetoma, paracoccidioidomycosis, melanomycosis, pseudoalescherichia,Examples include bacteria that cause trichorrhizoa, Pneumocystis, human metapneumovirus, human rhinovirus, enterovirus, influenza A, influenza B, Middle East respiratory coronavirus (MERS-CoV), parainfluenza virus 1, parainfluenza virus 2, parainfluenza virus 3, parainfluenza virus 4, syncytial viruses of the respiratory system, Bordetella pertussis, Bordetella pertussis, Chlamydia pneumonia, Mycoplasma pneumonia, and combinations thereof.

[0037] Suitable chemical substances include ketones, nicotine, cocaine, opioids, marijuana, benzodiazepines, amphetamines, and barbiturates.

[0038] The sample can also be analyzed for proteins, DNA, and RNA.

[0039] The sample can be analyzed simultaneously for multiple different analytes (e.g., multiplex assay). For example, the method of this disclosure is particularly suitable for detecting and identifying RNA from SARS-CoV-2, influenza A virus, and influenza B virus in upper or lower respiratory tract samples in multiplex assays.

[0040] Any method known in the art for analyzing the samples of this disclosure is suitable for analyzing bioaerosol samples collected using a bioaerosol collection device, as well as composite samples collected using a bioaerosol collection device, swabs, washes, aspirates, and combinations thereof. Suitable methods include, for example, polymerase chain reactions and other DNA and RNA amplification methods (e.g., PCR, RT-PCR, loop-mediated isothermal amplification (LAMP)), immunoassay detection methods (e.g., Western blot analysis and enzyme immunosorbent assay (ELISA)), chromatography (e.g., HPLC), gas chromatography, capillary pherosis, 2D and 3D gel electrophoresis, mass spectrometry, and combinations thereof.

[0041] The sample can be analyzed using a device such as a nitrocellulose lateral flow strip containing one or more capture zones: a control line to detect the presence of all antibodies in the sample, and a test line that specifically reacts with the analyte to be detected. The sample can also be analyzed using an antigen test. As is known in the art, an antigen test includes an antibody on a test device that binds to an antigen contained in the sample. In another embodiment, a combined sample can be analyzed with an antibody test. As is known in the art, an antibody test includes an antigen on a test device that binds to an antibody contained in the sample. In both antigen-antibody and antibody tests, the binding of antigen to antibody results in a visible result indicating that the subject is infected.

[0042] Typical lateral flow analysis ("LFA") and vertical flow analysis ("VFA") test strips include a double membrane mounted on a backing card for better stability and handling. The sample is applied to one end of the strip on an adsorption sample pad impregnated with buffer salts and surfactants to make the sample suitable for interaction with the detection system. The sample moves through a conjugate release pad containing antibodies that are specific to the target analyte and conjugated to colored or fluorescent particles. The sample, along with the conjugated antibody bound to the target analyte, moves along the strip into detection zones that have specific biological components (e.g., antibodies or antigens) immobilized in lines that react with the analyte bound to the conjugated antibody. Recognition of the sample analyte results in a response on the test line, while a response on the control line indicates a proper liquid flow through the strip. Readouts, represented by lines appearing at different intensities, can be evaluated visually or using a dedicated reader. Further test lines of antibodies specific to different analytes can be immobilized in array form to test multiple analytes simultaneously under the same conditions.

[0043] The analysis can be automated using commercially available platforms such as Cobas Amplicor (Roche Molecular Diagnostics, Pleasanton, California).

[0044] The analysis can utilize a point-of-care platform and / or an off-site high-throughput platform.

[0045] The bioaerosol collection device of this disclosure enables the collection of bioaerosol samples provided or acquired from a subject. The subject may direct the air sample through the mouth (oral), through one or both nostrils, and through a combination of mouth and nose. The subject directs the air sample (by blowing, exhaling, breathing, humming, singing, talking, counting, coughing, inhaling through the nose, and a combination thereof). The subject may expel air multiple times, for set periods, and in combination thereof. The air expelled by the subject enters the bioaerosol sample collection device through an inlet. The air moves to a capture substrate located within the bioaerosol collection device, where the analytes contained in the bioaerosol sample are captured. The bioaerosol collection device also includes an outlet (such as a vent) that allows the air to exit the bioaerosol collection device after passing through the capture substrate. The outlet is configured to reduce the pressure within the device so that the position of the capture substrate is not obstructed / disturbed during the bioaerosol collection process.

[0046] In one embodiment, the present disclosure relates to a bioaerosol collection device for collecting a bioaerosol sample. The bioaerosol collection device includes a hollow housing including an inlet, an outlet, and a passage that fluidly couples the inlet and the outlet, and a capture substrate positioned in the passage downstream of the inlet toward the outlet, wherein the bioaerosol sample comes into contact with the capture substrate as the bioaerosol sample flows from the inlet toward the outlet.

[0047] In one embodiment, the disclosure relates to a bioaerosol collection device for collecting a bioaerosol sample. The bioaerosol collection device comprises a hollow housing including an inlet, an outlet coupled to an analytical device, and a passage fluidly coupling the inlet and the outlet, and a capture substrate disposed in the passage downstream of the inlet, wherein the bioaerosol sample comes into contact with the capture substrate as it flows from the inlet to the outlet and extends through the outlet, with at least a portion of the capture substrate in contact with the sample area of ​​the analytical device.

[0048] As shown in Figure 1, the capture substrate is positioned between the inlet and the outlet. In certain embodiments, the capture substrate extends through the outlet (see, for example, Figure 14C).

[0049] Suitable capture substrates include those described herein.

[0050] As further described herein, the device is modular, meaning that elements can be attached to and detached from the device (see, for example, Figures 9 and 13). For example, the housing of the device may be a single unit or multiple units. Each housing unit may contain one or more components for sample collection, sample processing, and sample analysis. In the multiple housing embodiment, the housings may be joined by threading, friction fitting, etc. The housings may also be coupled to caps and adapters to connect the device to sample preparation and analysis devices, as described herein. In an exemplary embodiment of the modular device, a hollow tubular portion may be reversibly coupled to a housing that holds a collection substrate.

[0051] A hollow housing can be cylindrical, tubular, rectangular, elliptical, or in other shapes that allow air directed into the housing through an inlet to pass over or through the collection substrate and exit the housing through an outlet. The term "hollow" is used, in its usual sense, to refer to an empty space or cavity inside the housing member. The empty space allows for a directional airflow from the inlet through the housing to the device's outlet.

[0052] The device housing can be fabricated from any material. Particularly suitable materials for forming the device housing are those useful for detecting analytes in a sample and compatible with any buffers and reagents to be captured on the collection substrate. Preferably, the housing is made of a transparent, translucent, or light-transmitting material so that the user can see inside the housing. The housing may include a window that allows the user to observe the assay results. The window may also allow the user to transmit the assay results. For example, the user can transmit the results by taking a photograph of the assay and sending the photographic image electronically / Bluetooth to a receiver. The assay results can also be associated with a scannable code, such as a barcode, which allows the user to scan the barcode and transmit the test results electronically / Bluetooth®.

[0053] In one embodiment, the inner wall surface of the hollow housing may be hydrophobic. The inner wall can be made hydrophobic by forming the hollow housing using a hydrophobic material. Additionally or alternatively, the inner wall can be made hydrophobic by coating the surface of the inner wall of the hollow housing with a hydrophobic substance. In another embodiment, the inner wall of the hollow housing may be hydrophilic. Additionally or alternatively, the inner wall can be made hydrophilic by coating the surface of the inner wall of the hollow housing with a hydrophilic substance. Additionally or alternatively, part of the inner wall may be hydrophobic and other parts of the inner wall may be hydrophilic. The surface of the inner wall can also be treated to reduce surface tension and allow for easier flow of reagents and other liquids along the inner wall.

[0054] In one embodiment, the inner wall surface is adapted to contain a reagent. The surface can be adapted to contain a reagent by applying a reagent-containing coating to the inner wall surface and drying the coating. The dried coating can be hydrated by adding buffer or water to the device, thereby rehydrating the reagent, releasing it from the inner wall surface, and allowing the reagent to interact with the collection substrate.

[0055] The bioaerosol collection device may further include a filter (referred to herein as a “band filter”), as shown in Figure 2. The band filter can capture or trap larger particles contained in the bioaerosol sample. The filter is positioned close to the inlet so that particles larger than the analyte of interest are captured within the filter material before reaching the collection substrate. A preferred filter has a pore size of approximately 200 μm. Thus, particles in airflow larger than 200 μm are captured by the filter, while particles in airflow smaller than 200 μm pore size may pass through the filter and come into contact with the collection substrate. While a 200 μm pore size filter is preferred, the filter pore size can be increased or decreased depending on the size of the analyte of interest that is to be captured on the collection substrate.

[0056] In one embodiment, the filter is inserted into the same housing as the capture substrate. In another embodiment, the filter may be contained within a housing coupled to the housing that houses the capture substrate.

[0057] The bioaerosol collection device may further include a first closure configured to close the inlet, as shown in Figure 3. The bioaerosol collection device may further include a second closure configured to close the outlet, as shown in Figure 3. Suitable closures include screw caps, snap caps, and pressure-fit caps. A suitable cap may also include a cap having a buffer pack. The buffer pack contains the buffer components and reagents described herein. When the cap is tightened or pressure is applied to the cap, the buffer pack can burst, releasing the buffer into the device. As shown in Figure 5, the closure may include an aperture (or opening) that allows the device to function as a pressure-operated dropper. The liquid (buffer) can be introduced using the cap having the buffer pack or from a buffer vial. The inlet of the device is then closed. In certain embodiments, the housing or part of the housing of the bioaerosol collection device is made of a deformable material (flexible polymer). The user can apply pressure to the deformable part of the housing to transfer droplets through the pressure-operated dropper cap. The droplets can be transferred to vials and / or analytical devices, and the remaining liquid can be transported and / or stored.

[0058] Referring to Figure 6, the bioaerosol collection device includes an air and reagent flow chamber having an inner diameter that guides or directs the air sample toward the capture substrate. In some embodiments, the end proximal to the inlet has an inner diameter D1 that is larger than the region distal to the inlet (proximal to the capture substrate) (inner diameter D2). This allows for concentration of the airflow as the bioaerosol sample reaches the capture substrate. In certain embodiments, the inner diameter of the device proximal to the capture substrate is larger than the inner diameter of the device proximal to the inlet (see, e.g., Figure 10). An outlet (e.g., a vent) allows the air that has flowed through the capture substrate to exit the device (see Figure 7). This reduces the pressure on the capture substrate that could move or displace it. A bridge material in contact with the capture substrate uses capillary action and / or microfluidic channels to transfer the buffer containing the analyte released from the capture substrate to the assay components, initiating the analyte detection process. The device may further include a window for visualizing the results of the analytical assay.

[0059] The bioaerosol collection device may further include a capture substrate support (see, for example, Figures 4 and 10). The capture substrate support is in contact with a portion of the capture substrate to maintain the position of the capture substrate within the device and / or resist air pressure during the collection of the bioaerosol sample (i.e., when the object guides air into the device). The support may also be a shelf (see, for example, Figure 4), a cone (or conical) shape (see, for example, Figure 10), a basket shape, and other shapes corresponding to the shape of the capture substrate. The capture substrate support may be a spoke system that holds the capture substrate open without causing a pressure drop.

[0060] As shown in Figure 8, the device may further include buffer and / or reagent components on the inner surface of the housing. These buffer / reagent components may be, for example, in a lyophilized form that is hydrated when the buffer is introduced.

[0061] The bioaerosol collection device may further include an analytical device reversibly coupled to the bioaerosol collection device (see, for example, Figures 16A and 16B). Suitable analytical devices are described herein. As shown in Figure 4, the analytical device (e.g., a lateral flow assay (LFA) / vertical flow assay (VFA)) is coupled to the device. As shown in Figures 10-12, the capture substrate extends through the outlet and comes into contact with components of the analytical device (e.g., the sample pad of the LFA / VFA). As shown in Figure 13, a buffer can be introduced into the device to initiate sample analysis. Generally, the buffer leads to the elution / extraction of the analyte from the capture substrate, and the analyte then flows into the analytical device where detection is performed.

[0062] In the exemplary embodiment shown in Figure 15, the bioaerosol collection device can be formed in a "T-shaped" design so that the bioaerosol sample first flows perpendicular to the capture substrate, thereby removing some air pressure on the capture substrate. Figure 15 also shows the use of a cap with a band filter and buffer pouch, as described herein, and / or the application of buffer / reagents to the inner surface. The conical capture substrate extends through the outlet and comes into contact with components of the analytical device (e.g., LFA / VFA).

[0063] As shown in Figures 16A and 16B, the device may be modular, thereby allowing the device components to be separated from each other. As shown in Figure 16A, for example, the housing and the capture region can be separated from the analytical device. As shown in Figure 16B, the housing can be separated from the part of the device that is coupled to the analytical device.

[0064] As further shown in Figure 17, the bioaerosol collection device can be closed using a closure such as a cap for storage and transport of the capture device after the bioaerosol sample has been collected. Figure 17 also shows that the buffer can be added directly from a buffer vial and / or using a cap with a buffer pack as described herein.

[0065] The modularity of the bioaerosol collection device is further shown in Figure 18. Figure 18 shows the use of a coupling mechanism, such as threading, between the upper ("air & reagent flow chamber") and lower chambers to hold the capture substrate in place within the device housing. As shown in Figures 19A–19C, the modular upper and lower chamber components of the bioaerosol collection device can be separated by using a torsional motion to orient the upper and lower chambers together and / or pull them apart.

[0066] As shown in Figure 20, the capture substrate can be transferred out of the bioaerosol capture device. Any method for removing the capture substrate is preferred. For example, as shown in Figure 20A, the capture substrate can be removed using a stick and pushed out through the exit (or inlet). As shown in Figure 20B, pressure can be applied to move the capture substrate by pressing an internal plunger device having a slide knob located on the outside of the housing. To prevent puncturing the capture substrate, the transfer stick may have a plunger that applies more dispersed pressure over the capture substrate. As shown in Figure 20C, the capture substrate may have perforations near its outer edge, allowing the capture substrate to be torn away from its outer edge and the majority of the capture substrate to be transferred out of the device.

[0067] As shown in Figures 21A and 21B, the capture substrate can be directly transferred into a vial that is reversibly coupled to the device. Figures 22–24 show exemplary embodiments of a bioaerosol collection device configured to reversibly couple to a vial.

[0068] As described herein, one aspect of the present disclosure relates to a bioaerosol collection device including an analytical device. Figure 25 shows a bioaerosol collection device having a cap and an integrated analytical device (e.g., LFA). When in use, the device directs an air sample towards the inlet. The air flows through an optional band filter and through a capture substrate that captures analytes contained in the air. When the inlet and outlet of the device are closed (not shown) and the cap is tightened, a buffer pack bursts and the buffer is released onto the capture substrate. The buffer transfers the analytes to a junction where analyte detection begins. A window allows the user (or another person) to view the analytical test results. As shown in Figure 26, the device may further include an opening that allows a swab to be introduced into the device. As shown in Figure 26, it is desirable that the swab collection material and the capture substrate be placed in close proximity so that when the buffer is released, the buffer comes into contact with both the swab and the capture substrate to obtain a mixed sample.

[0069] In one embodiment, the present disclosure relates to a bioaerosol collection device for collecting a bioaerosol sample. The bioaerosol collection device comprises a capture substrate configured to capture an analyte in a bioaerosol sample; a first bioaerosol permeable protective layer disposed on the upper surface of the capture substrate and covering the upper surface of the capture substrate; and a second bioaerosol permeable protective layer disposed on the lower surface of the capture substrate and covering the lower surface of the capture substrate.

[0070] As shown in Figure 27, the bioaerosol collection device has an upper outer layer, a capture substrate layer, and a lower outer layer. The capture substrate layer includes a capture substrate. As shown in Figure 27, air flows through each layer. Analytes in the bioaerosol sample are captured by a capture substrate as described herein. Suitable capture substrates are described herein. As shown in Figures 27 to 37, each component of the bioaerosol collection device together forms a passage. The capture substrate is placed within the passage, thereby bringing the bioaerosol sample into contact with the capture substrate as the bioaerosol sample flows. The user directs the bioaerosol sample into the capture area, and the bioaerosol sample flows through the upper outer layer, through the capture substrate, and through the lower outer layer. The capture areas of the upper and lower outer layers may include protective materials (e.g., filters).

[0071] In certain embodiments, the bioaerosol collection device further includes a holder coupled to a capture substrate (shown in Figure 27). In some embodiments, the holder is releasably coupled to the capture substrate. The holder is coupled to the capture substrate using preferred fasteners (e.g., pins, rivets, screws, compression pins, etc.), adhesives, ultrasonic welding, and combinations thereof.

[0072] The bioaerosol collection device may further include a support layer having an opening of a size and shape corresponding to the size and shape of the capture substrate. It should be understood that the opening and shape of the support layer do not need to be exactly the same size and shape as the capture substrate. For example, the size and shape of the capture substrate can be slightly larger than the size and shape of the opening in the support layer, so that the outer edge of the capture substrate overlaps with the edge of the opening in the support layer, providing a contact surface between the capture substrate and the support. The size and shape of the capture substrate can be slightly smaller than the size and shape of the opening in the support layer so that the capture layer "floats" within the opening in the support layer.

[0073] The support layer may further include a holder coupled to the capture substrate, thereby the support being releasably connected to the holder, and the holder being configured to be detachable from the support in order to remove the capture substrate from the rest of the bioaerosol collection device.

[0074] As shown in Figure 28, the bioaerosol collection device may further include an airflow ring (also referred to herein as an "airflow tube"). The airflow ring / tube instructs the user where to position their lips, mouth, nostrils, or nostrils when preparing to provide a bioaerosol sample. The airflow ring / tube can be made from any suitable material. The airflow ring / tube may be made from a soft foam to provide comfort and help seal the user's skin surface to the airflow ring / tube.

[0075] As shown in Figures 28 and 32-37, the bioaerosol collection device may further include an upper protective layer and a lower protective layer. The protective layer(s) can capture (i.e., filter and remove) undesirable particles contained in the bioaerosol sample, prevent contamination of the capture substrate by accidental contact with the capture substrate, and prevent physical damage to the capture substrate.

[0076] As shown in Figure 29, the outer layer may include folds to facilitate removal of the outer layer when it is desirable to expose the capture substrate (for example, to process and test the capture substrate). The outer layer can be peeled off like an adhesive bandage. Figure 29 also shows the use of an adhesive line proximal to the capture area to provide additional resistance while the outer layer is separated.

[0077] As shown in Figure 30, a portion of the capture substrate holder may extend beyond the outer layer. The user can hold the exposed handle while removing the outer layer (one or both layers). In some embodiments, the handle of the capture substrate holder includes a break point that allows the handle to be broken into a shorter length. This allows the handle to be shortened so that it can be placed inside the vial.

[0078] As shown in Figure 31, the outer layer can be slightly larger in size and / or shape than the capture substrate and / or protective layer. This allows for the use of less material while providing support and protection for the capture substrate. Figure 31 also shows that the edges of the device that come into contact with the user's skin surface can be flat or arc-shaped, which can create a better seal with the user's skin surface and / or guide the user when positioning the device for bioaerosol sample collection.

[0079] Figure 32 shows one embodiment including a holder to which a capture substrate layer is bonded to a capture substrate. Figure 33 shows one embodiment in which the capture substrate layer includes a support layer with an opening of a size and shape corresponding to the size and shape of the capture substrate, the support is releasably connected to the holder, and the holder is configured to be detached from the support to remove the capture substrate from the rest of the bioaerosol collection device. In some embodiments, the holder is releasably bonded to a capture substrate as described herein. The holder can be detached from the support material.

[0080] The support and the substrate capture holder can be semi-rigid or rigid. Suitable semi-rigid and rigid materials are known in the art, such as plastics, polymers, and metals. High-density polyethylene is a particularly suitable material for the support and the substrate capture holder.

[0081] As shown in Figure 35, the holder can be reversibly bonded to the support using tabs made of adhesive material or similar. The use of an upper tab and at least one lower tab allows for reversible bonding of the capture substrate holder to maintain the position of the capture substrate and the capture substrate holder. The tabs may also include writing for providing instructions for use or other information.

[0082] Figure 37 shows a bioaerosol collection device having an arc-shaped edge similar to the exemplary embodiment shown in Figure 31, which can form a better seal with the user's skin surface and / or guide the user when positioning the device for bioaerosol sample collection.

[0083] Figure 38 shows an exemplary embodiment of the capture layer. The capture layer may be a capture substrate alone. The capture substrate may also include perforations and / or adhesives as described herein. The capture substrate may also include slits (as shown in Figure 38), folds and creases to increase the surface area of ​​the capture substrate and enable the capture substrate to be used as a swab. The capture substrate may be connected to a holder that can be removed from a support. As also described herein, the capture substrate may be permanently or reversibly bonded to a holder as described herein.

[0084] Figure 39 shows an exemplary embodiment of a capture holder. As shown in Figure 39, the capture substrate layer includes a support that is releasably connected to the holder, and the holder is configured to be separable from the support. The holder can be bonded to the capture substrate as described, using adhesive, welding, fasteners, etc. The capture substrate holder can be sized and shaped to support the outer edge of the capture substrate and may include a support network to support the middle portion of the capture substrate. Also, as described and illustrated in Figure 39, the holder includes a break point that allows the length of the holder to be reduced (shortened).

[0085] In one embodiment, the present disclosure relates to a bioaerosol collection device for collecting a bioaerosol sample. The bioaerosol collection device comprises a capture substrate configured to capture an analyte in a bioaerosol sample; a first compartment configured to cover the upper surface of the capture substrate; and a second compartment configured to cover the lower surface of the capture substrate and comprising an analytical device, the analytical device comprising a sample pad that is in fluid contact with the lower surface of the capture substrate. The first and second compartments are configured to be joined together to form a seal. When a buffer solution is introduced onto the capture substrate, the analyte is released from the capture substrate into the buffer solution, and the buffer solution flows to the sample pad of the analytical device.

[0086] As shown in Figure 40, the upper and lower compartments are in the form of a “clamshell” enclosing the capture substrate. When closed, the upper and lower compartments form a fluid-sealed seal. In certain embodiments, the upper and lower compartments can be joined using hinges. In certain embodiments, the upper and lower compartments exist as separate components. Figure 40 also shows an exemplary embodiment in which the upper compartment includes a buffer pack described herein, which releases a buffer when pressure is applied to join the upper and lower compartments. Part of the capture substrate holder can also contact the buffer pack to assist in the release of the buffer. The lower compartment may include additional features that contact the buffer pack to assist in the release of the buffer when the upper and lower compartments are closed. In some embodiments, the lower compartment includes an assay device such as an LFA / VFA. A bridging material can contact the smaller end of the capture substrate and the sample pad of the VFA / LFA to transfer the sample to the LFA / VFA. Figure 41 shows the device in a closed configuration in top and bottom views of the device. Figure 42 shows an exemplary embodiment of the device including a support and tabs. Figure 43 shows an exemplary embodiment of the device in which the upper compartment includes a well that allows buffer to be introduced into the capture substrate through the upper compartment.

[0087] Figures 44 and 45 are photographic images of an exemplary embodiment of a bioaerosol collection device.

[0088] Figure 46 illustrates an exemplary embodiment of a punch-design bioaerosol collection device. The pore diameter of the lower outer layer can be smaller than that of the upper outer layer. The upper pore diameter can accommodate the shape of the patient's mouth and / or nostrils, and the lower pore diameter can be smaller than the diameter of the vial through which the internal collection medium is transported. The capture substrate is either fully perforated or frictionally held within the device. The lower and upper protective layers are almost fully perforated. When pressure is applied, the upper and lower protective layers detach but remain attached to the outer layer. The user can push down the upper outer layer through the collection area to fully move and release the capture substrate and transport it into the vial. The upper and lower outer layers and protective layers may remain hinged to the device.

[0089] Figure 47 illustrates an exemplary embodiment of a bioaerosol collection device having a cap system. The upper and lower layers have threaded or friction points that engage with the cap. When a user is introducing a bioaerosol sample, an optional protective layer for one or both of the upper and lower layers can be placed on the capture substrate. Figure 47 also shows that writing may be applied to the device.

[0090] Figures 48 and 49 illustrate exemplary embodiments of a bioaerosol collection device having an airflow tube adapter. The airflow adapter includes an insertion opening for receiving the bioaerosol collection device. For comfort, any soft material may be included on the adapter. This device is particularly useful for collecting bioaerosol samples from multiple users using the same bioaerosol collection device. When in use, each user has their own adapter, but a single bioaerosol capture device is used by all users. For example, a family of five can blow into the same sample collection device being tested. Figure 49 shows an airflow adapter configured for both nostrils of a user.

[0091] In the exemplary embodiment shown in Figure 50, the bioaerosol collection device includes a buffer pack, a channel for fluidly coupling the buffer pack to a capture substrate, and a second channel for fluidly coupling the capture substrate to an analytical device. Figure 51 shows an exemplary bioaerosol collection device coupled to an analytical device (LFA / VFA) and transferred to a vial containing reagent buffer that contacts the capture substrate to initiate analyte detection.

[0092] In another aspect, the Disclosure relates to a method for collecting and analyzing a bioaerosol sample, comprising: obtaining a bioaerosol sample from a subject using a bioaerosol collection device; obtaining a swab sample from the subject; combining the bioaerosol sample and the swab sample; and analyzing the combined sample.

[0093] As shown in Figures 52 and 53, the user directs the bioaerosol sample into the bioaerosol collection device. The bioaerosol sample can be provided through the user's mouth or nasal cavity. Swabs are also used to collect nasopharyngeal (or oral) samples. As shown in Figure 52, the swab sample is eluted using a suitable buffer solution, and the swab eluate is then introduced into the bioaerosol collection device to elute the analyte from the capture substrate. The combined eluate flows into the analytical device for analysis. As shown in Figure 53B, the swab can be inserted into the bioaerosol collection device. The buffer solution can be added from a buffer vial and / or via a cap system having a buffer pack as described herein. The proximity of the swab and the capture substrate allows for sample combination, and the sample flows to the junction with the analytical device for analysis.

[0094] A suitable bioaerosol collection device is described herein.

[0095] As shown in Figure 54, bioaerosol and swab samples are collected from the subject. Elution / extraction buffer is added to the dropper vial. Using a swab, the captured substrate is transferred from the bioaerosol collection device to the dropper vial containing the elution / extraction buffer. The combined sample droplets are then introduced into the analytical device.

[0096] In another embodiment illustrated in Figure 55, the bioaerosol capture substrate and swab are eluted in separate vials of extraction / elution buffer and then combined to form a composite sample. In another embodiment shown in Figure 56, the bioaerosol capture substrate and swab are eluted in the same vial of extraction / elution buffer to form a composite sample. In another embodiment illustrated in Figure 57, the swab is eluted in a vial of extraction / elution buffer and then discarded. The swab eluate is then used to elute / extract the bioaerosol capture substrate to form a composite sample.

[0097] The method of this disclosure combines a sample obtained using a bioaerosol collection device. In another embodiment, the method of this disclosure can combine the bioaerosol sample and the first sample with a third sample collection method. Suitable third samples include, for example, sputum, nasal irrigation solution, oral rinse solution, and oral swabs.

[0098] In another embodiment, the bioaerosol sample eluate can be stored and / or transported for analysis in a laboratory testing facility. After collection, the sample can be placed in a sterile transport container containing a transport medium. Suitable transport media include, for example, viral transport medium (VTM), amies transport medium, and sterile saline. The transport medium may include, for example, RNase inhibitors, DNase inhibitors, protease inhibitors, preservatives, stabilizers, and other reagents for preserving the sample. Once placed in the transport container, the collected and combined sample can be stored for subsequent analysis or processed for analysis. Processing may include dissolution, extraction, and other known processing steps for analyzing the mixed sample for analytes.

[0099] Any method known in the art for analyzing the samples of this disclosure is suitable for analyzing bioaerosol samples collected using a bioaerosol collection device, as well as composite samples collected using a bioaerosol collection device, swabs, washes, aspirates, and combinations thereof. Suitable methods include, for example, polymerase chain reactions and other DNA and RNA amplification methods (e.g., PCR, RT-PCR, loop-mediated isothermal amplification (LAMP)), immunoassay detection methods (e.g., Western blot analysis and enzyme immunosorbent assay (ELISA)), chromatography (e.g., HPLC), gas chromatography, capillary pherosis, 2D and 3D gel electrophoresis, mass spectrometry, and combinations thereof. [Examples]

[0100] Example 1 This embodiment outlines the testing of different sample collection methods and combinations of sample collection methods.

[0101] 1. Device: Blow tube

[0102] Use case: general population

[0103] Detection route: PCR for multi-site (oral + NP) swabs vs. PCR for bioaerosols vs. sputum.

[0104] Pathogen tested: COVID+TB (a single-elution protocol needs to be determined)

[0105] Registration criteria: Presumed positive

[0106] Scientific necessity: To compare rapid bioaerosol collection and NP swab collection methods from 2 days to 7 days before the onset of COVID symptoms. Sputum for TB

[0107] Behavior regarding bioaerosol collection: Counts up to 20 deep breaths and 10 coughs.

[0108] Collection time: 10-15 minutes

[0109] Sample handling: The POC operator removes the bioaerosol sample collector from the device and places it in a 5 mL Eppendorf 1 mL UTM. The sample is obtained in buffer by mechanical vibration for 10 seconds and rotation for 30 minutes.

[0110] 2. Device: Blow tube

[0111] Use case: general population

[0112] Detection route: PCR for multi-site (oral + NP) swabs vs. PCR for bioaerosols

[0113] Pathogens tested: Respiratory panel

[0114] Registration criteria: Cough, shortness of breath, or pharyngitis of any duration

[0115] Scientific necessity: For respiratory panel diseases, compare rapid bioaerosol collection versus NP swab collection methods, ideally from 2 days to 7 days before symptom onset.

[0116] Behavior regarding bioaerosol collection: Counts up to 20 deep breaths and 10 coughs.

[0117] Collection time: 10-15 minutes

[0118] Sample handling: The POC operator removes the bioaerosol sample collector from the device and places it in a 5 mL Eppendorf 1 mL UTM. The sample is obtained in buffer by mechanical vibration for 10 seconds and rotation for 30 minutes.

[0119] 3. Device: Blow tube

[0120] Use case: general population

[0121] Detection route: Multi-site (oral + NS) swabs + PCR for bioaerosol vs. PCR for NP swabs

[0122] Pathogens tested: COVID+TB

[0123] Registration criteria: Presumed positive

[0124] Scientific necessity: To compare rapid bioaerosol collection combinations with NP swab collection methods from 2 days to 7 days before the onset of COVID symptoms.

[0125] Actions for bioaerosol collection: Deep breaths x 20, coughs x 10, count up to 20.

[0126] Collection time: 10-15 minutes

[0127] Sample handling: The POC operator removes the bioaerosol sample collector from the mask and places it in a 5 mL Eppendorf 1 mL UTM. Obtain the sample in buffer by mechanically vibrating for 10 seconds and rotating for 30 minutes.

[0128] 4. Device: Blow tube

[0129] Use case: general population

[0130] Detection route: Multi-site (oral + NS) swabs + PCR for bioaerosol vs. PCR for NP swabs

[0131] Pathogens tested: Respiratory panel

[0132] Registration criteria: Cough, shortness of breath, or pharyngitis of any duration

[0133] Scientific necessity: For respiratory panels, we will compare rapid bioaerosol sample combinations and NP swab collection methods from 2 days before symptom onset to 7 days later.

[0134] Actions for bioaerosol collection: Deep breaths x 20, coughs x 10, count up to 20.

[0135] Collection time: 10-15 minutes

[0136] Sample handling: The POC operator removes the bioaerosol sample collector from the mask and places it in a 5 mL Eppendorf 1 mL UTM. Obtain the sample in buffer by mechanically vibrating for 10 seconds and rotating for 30 minutes.

[0137] 5. Device: Blow tube attached to the RDT

[0138] Use case: general population

[0139] Detection route: Multi-site (oral + NP) swab by PCR vs. bioaerosol by RDT (visual and reader-based).

[0140] Pathogen tested: COVID

[0141] Registration criteria: Presumed positive

[0142] Scientific necessity: Ideally, compare qualitative testing by RDT using only rapid bioaerosol samples from 2 days to 7 days before symptom onset in a respiratory panel with quantitative NP swab collection tested by reference PCR.

[0143] Actions for bioaerosol collection: Deep breaths x 20, coughs x 10, count up to 20.

[0144] Collection time: 10-15 minutes

[0145] Sample handling: The POC operator attaches the bioaerosol collector to the RDT. The patient performs the procedure. The POC applies the required elution / extraction buffer droplets to the bioaerosol substrate.

[0146] 6. Device: Blow tube attached to the RDT

[0147] Use case: general population

[0148] Detection route: Multi-site (oral + NP) swabs by PCR vs. NS swabs + bioaerosol by RDT (visual and reader-based detection).

[0149] Pathogen tested: COVID

[0150] Registration criteria: Presumed positive

[0151] Scientific necessity: Ideally, compare qualitative testing of rapid bioaerosol samples combined with NS samples via RDT from 2 days to 7 days before symptom onset in a respiratory panel, with quantitative NP swab collection tested via reference PCR.

[0152] Actions for bioaerosol collection: Deep breaths x 20, coughs x 10, count up to 20.

[0153] Collection time: 10-15 minutes

[0154] Sample Handling: The POC operator attaches the bioaerosol collector to the RDT. The patient performs the bioaerosol collection behavior within the device. The POC operator swabs the patient with an NS swab and elutes into the RDT elution / extraction buffer as usual. The POC applies the required elution / extraction buffer droplets to the bioaerosol substrate and tests via the RDT.

[0155] The embodiments described herein involve combining a sample obtained using a bioaerosol collection device with a sample collected using a swab collection method, but it should be understood that the sample obtained using the bioaerosol collection device can be analyzed alone (without being combined with the second sample). Advantageously, the method of the disclosure makes the viral load available at a density that can be used in a wide variety of tests, such as amplification, antibody, antigen, and other paper-based tests. When the method of the disclosure combines a sample collected using a bioaerosol collection device with a swab sample, the sample combination is particularly advantageous for specifically increasing the availability of the viral load.

[0156] Example 2 In this example, nasal air samples collected using an exemplary embodiment of a sample collection device were analyzed for the presence of COVID.

[0157] Air samples were collected from patients suspected of being COVID-positive. Patients also blew air samples into a collection device through their nostrils (samples labeled "S" from patient ID). Nasal swab samples were also collected and analyzed from each patient (samples labeled "B" from patient ID).

[0158] Analytes collected on the device's collection substrate were eluted with a buffer containing TX45. PCR was performed on each sample type using the TaqPath kit to detect MS2, N gene, ORF1ab, and S gene.

[0159] As summarized in the table in Figure 59, positive detection was obtained from nasal air samples. Surprisingly, the detection in nasal air samples was nearly 100% consistent with that of samples collected using nasopharyngeal swabs.

[0160] This embodiment demonstrates that a device can be used to collect nasal air samples and detect analytes contained in the nasal air samples.

[0161] The methods of this disclosure advantageously enable the overall sensitivity of the test and reduce false negative test results. In particular, combining exhaled droplet samples with swab samples can maximize test sensitivity and reduce the amount of test resources used to analyze the samples. Furthermore, combining bioaerosol samples and released viruses using a bioaerosol collection device with other samples collected using different collection methods advantageously increases the amount of collected virus that can be tested, thereby increasing test sensitivity. Combining bioaerosol samples and released viruses collected using a bioaerosol collection device with samples collected using a swab collection method can also advantageously smooth out signal attenuation inherent in upper respiratory tract sampling for saliva samples, for example. Combining a bioaerosol collection device with samples collected using swab collection can also advantageously assist downstream test handling. Combining a bioaerosol collection device with samples collected using swab collection can also advantageously reduce false negatives because it can increase the amount of pathogens collected using at least two different sample collection methods.

Claims

1. A bioaerosol collection device for collecting bioaerosol samples, It is a hollow housing, The entrance and Exit and A hollow housing including a passage that fluidly connects the inlet and the outlet, A bioaerosol collection device comprising: a capture substrate including an electret, which is disposed in the passage downstream of the inlet and configured to have a lower surface for capturing analytes in the bioaerosol sample, wherein the lower surface is positioned to face the outlet side.

2. The bioaerosol collection device according to claim 1, wherein the capture substrate is disposed between the inlet and the outlet.

3. The bioaerosol collection device according to claim 1, wherein the capture substrate extends through the outlet.

4. The bioaerosol collection device according to claim 1, further comprising a first closure configured to close the inlet.

5. The bioaerosol collection device according to claim 1, further comprising a second closure configured to close the outlet.

6. The bioaerosol collection device according to claim 1, further comprising an analytical device reversibly coupled to the housing.

7. A bioaerosol collection device for collecting bioaerosol samples, It is a hollow housing, The entrance and An outlet coupled to the analysis device, A hollow housing including a passage that fluidly connects the inlet and the outlet, A bioaerosol collection device comprising a capture substrate disposed in the passage downstream of the inlet and configured to capture analytes in the bioaerosol sample, wherein at least a portion of the capture substrate is in contact with the sample area of ​​the analysis device.

8. The bioaerosol collection and analysis device according to claim 7, wherein the capture substrate has a conical shape including a tip that contacts the sample area of ​​the analysis device.

Images