Compositions, devices, and methods for testing inanimate surfaces, populations, and individuals for pathogen infection

Abstract

Disclosed herein are compositions, devices, and methods for detecting analytes in an air sample taken from a subject. The compositions can comprise on an inanimate surface, such as a face mask worn by an individual, having a test substrate for capturing analytes.

Description

[0001] Cross-references to related applications

[0002] This application claims priority and benefit to U.S. Provisional Patent Application Serial No. 63 / 029,974, filed May 26, 2020, which is incorporated herein by reference in its entirety. Background Technology

[0003] This disclosure generally relates to medicine, as well as industry and environmental health in general. More specifically, this disclosure relates to compositions, apparatus, and methods for using analytes in air samples obtained from individuals.

[0004] Medical monitoring and diagnosis involve sample collection and analysis. Sample collection can involve invasive and non-invasive methods. Invasive methods of sample collection include procedures such as surgery and blood draws, which can cause pain, discomfort, and stress for the patient. Milder forms of invasive sample collection may include swabbing. The devices used in invasive sample collection methods can also be expensive and require sterilization to prevent sample contamination. Non-invasive sample collection methods may be superior to invasive methods in reducing pain and discomfort for individuals during sample collection.

[0005] Rapid detection and identification of pathogens are crucial for preventing and monitoring the presence and spread of infections, including preventing and mitigating the spread of global pandemics. Testing surfaces and individuals to detect the presence of pathogens allows for appropriate diagnosis, further testing, and / or treatment of individuals. Individuals can also be tested to detect circulating antibodies against pathogens, indicating that the individual has been exposed to the pathogen in the past and has developed an immune response against the infection.

[0006] The detection and identification of analytes, such as biomarkers, can aid in the diagnosis and / or progression of diseases, determine the effectiveness and / or response to treatment, and allow for the correction and / or adjustment of treatment dosages. Analyte detection can also be used to detect environmental exposure to potentially harmful chemicals and drug use (such as illicit drug use). Biomarker detection can also provide information related to internal physiological states, including disease, metabolic status, toxins, and the presence of certain chemicals.

[0007] Current testing is insufficient to meet societal needs during pandemics such as the COVID-19 pandemic. For example, at one point during the COVID-19 pandemic, approximately 500,000 tests were being conducted daily, while more than 20 million tests were needed daily, highlighting a gap in testing scale. Current testing also faces shortcomings and challenges, such as scalability, sample collection and extraction, testing costs, pain associated with sample collection, transmission risks, test invalidity (e.g., temperature checks), and failure to test asymptomatic individuals. Current testing also misses essential metrics about who is spreading the disease.

[0008] Current testing methods using nasal swabs for sample collection are hampered by limited swab supply, sample collection can be painful and uncomfortable, and requires collection by trained healthcare professionals. Sample collection (such as saliva, sputum, saliva, irrigating fluid, and other oral fluid samples) may require specialized equipment and further processing to separate analytes from the oral fluid. Regardless of the sample collection method used, the acquired sample may begin to degrade, and sample collection reagents may be affected by supply chain delays. The sample must subsequently be removed from or separated from the collection device, which can be inefficient and affect the sensitivity of the tests used to analyze the sample. Collection devices for saliva and nasal samples add complexity and are susceptible to contamination by the underlying sample collection matrix; i.e., saliva and nasal swabs contain numerous chemicals that need to be removed before extracting the target analyte. Proteases and ribonucleases can also degrade the target analyte after extraction. The ratio of the surface area of ​​the collection material to the necessary elution and extraction buffers affects the concentration of the target analyte. Higher concentrations of the target analyte result in more sensitive tests. Optimal material selection for shape factor and performance provides maximum test sensitivity.

[0009] Therefore, there is a need to develop compositions, devices comprising these compositions, and methods for using these compositions for analyte testing on surfaces, populations, and individuals. Low-cost testing of exhaled bioaerosols globally remains a critical unmet market demand.

[0010] Brief description

[0011] On one hand, this disclosure relates to a mask insert comprising a first layer, a test substrate, and a second layer, the test substrate being used to capture analytes in an air sample obtained from a subject, wherein the first and second layers form an outer layer substantially surrounding the test substrate, wherein overlapping areas of the first and second layers are combined, wherein at least a portion of the test substrate is combined with at least one of the first and second layers, and wherein the test substrate is configured to be separated from at least a portion of the outer layer.

[0012] On one hand, this disclosure relates to a system comprising: a mask insert including a first layer, a test substrate, and a second layer, the test substrate being used to capture analytes from an air sample obtained from a subject, wherein the first and second layers form an outer layer substantially surrounding the test substrate, wherein overlapping areas of the first and second layers are combined, wherein at least a portion of the test substrate is combined with at least one of the first and second layers, and wherein the test substrate is configured to be detached from at least a portion of the outer layer; and a mask worn by a user, wherein the mask insert is attached to the surface of the mask.

[0013] On the other hand, this disclosure relates to a method for detecting the presence of analytes in an air sample obtained from a subject, the method comprising: collecting a test substrate worn by the subject from an individual; and analyzing the analytes on the test substrate. Attached Figure Description

[0014] This disclosure will be better understood when taken into consideration its following detailed description, and features, aspects, and advantages other than those set forth above will become apparent. This detailed description refers to the following figures, in which:

[0015] Figure 1 This is an illustration depicting a front view of one embodiment of a mask insert.

[0016] Figure 2 This is an illustration of a front view depicting one embodiment of the mask insert, showing partial separation of the outer layer to expose the test substrate. The patient or healthcare provider clamps the outer layer at each end of the mask insert and pulls it outward in the direction of the large arrows (the pulling force is indicated by the large arrows). The clamping points of the device can be asymmetrical. The clamping point on the right side of the illustration clamps both the outer layer and the inner test substrate. The clamping point on the left side of the illustration clamps only the outer layer. By clamping the ends and pulling outward, the perforations allow removal of the outer protective layer on the left side while retaining the protective layer to secure the inner test substrate on the right side. After partial removal of the outer layer, the patient or healthcare provider secures the outer protective layer at the clamping point on the right side of the illustration to protect the test substrate from contact with hands that may contain ribonuclease, protease, and other contaminants. The patient or healthcare provider can then place the collected substrate in a vial for testing.

[0017] Figure 3 These are illustrations depicting the front and side views of a mask insert. The side view illustrates the layering and positioning of the micro-hooks used to attach the mask insert to the mask.

[0018] Figure 4 This is an illustration depicting a mask insert with a buffer orifice and an integrated vertical flow measurement unit.

[0019] Figure 5This is an illustration depicting a mask insert with a buffer dispensing device and an integrated vertical flow measurement unit.

[0020] Figure 6 This illustration depicts a mask insert (outer layer removed) with a buffer dispensing device and an integrated vertical flow measurement unit. A multi-stage vertical flow measurement platform is also depicted.

[0021] Figure 7 This is an illustration depicting a mask insert (outer layer removed) with three buffer dispensing devices and an integrated vertical flow multiplexing unit.

[0022] Figure 8 This is an illustration depicting a mask insert (outer layer removed) with three buffer dispensing devices and an integrated vertical flow multiplexing unit.

[0023] Figure 9A This is an illustration depicting a mask insert (outer layer removed) with three buffer dispensing plungers and an integrated vertical flow multiplexing unit.

[0024] Figure 9B This is an enlarged diagram depicting a three-buffer solution dispensing plunger assembly.

[0025] Figure 10 This is an illustration depicting a mask insert (outer layer in proper position) with a buffer orifice and an integrated sideflow measuring unit.

[0026] Figure 11 This is an illustration depicting a mask insert (outer layer in proper position) with a buffer dispensing device and an integrated sideflow measuring unit.

[0027] Figure 12 This is an illustration depicting a mask insert (outer layer removed) with a buffer dispensing device and an integrated sideflow measuring unit.

[0028] Figures 13A to 13C An exemplary embodiment of the test substrate is depicted, showing the splice. Figure 13A An exemplary embodiment of a test substrate is depicted, having four tab portions and a label portion. The test substrate may have perforations parallel to the top edge (and located below the label portion) and positioned between the individual tabs, allowing the tabs to be removed independently from the test substrate. Figure 13B An exemplary embodiment of a test substrate is depicted, having four tab portions and a label portion. These tab portions are spaced apart but attached to the label portion. The test substrate may have perforations parallel to the top edge (and located below the label portion) and positioned between the individual tabs, allowing the tabs to be removed independently from the test substrate. Figure 13C An exemplary embodiment of a test substrate is depicted, which has four unconnected tab portions.

[0029] Figure 14 The embodiment of Figure 13 is depicted as part of a face mask worn by a user.

[0030] Figure 15 An exemplary embodiment of a multilayer test substrate is depicted, having four tab portions and an inner test substrate that is protected on all sides and is made of a material with a high surface area that can be released from the outer protective layer.

[0031] Figure 16 A flowchart for extracting and amplifying RNA from the test substrate is depicted.

[0032] manual

[0033] This disclosure generally relates to compositions, apparatus, and methods for testing pathogen infection on inanimate surfaces, in groups, and in individuals. In particular, this disclosure relates to apparatus for detecting analytes in air samples exhaled by a subject.

[0034] In one respect, this disclosure relates to a mask insert. (See reference) Figures 1 to 3 The mask insert includes a first outer layer, a test substrate, and a second outer layer. One of the first or second layers faces (close to) the subject's face, while the other layer is attached (attached) to the inner surface of the mask worn by the subject.

[0035] like Figures 1 to 3 As shown, the first and second layers are wider and longer than the test substrate (marked with hash symbols and their outer boundaries are dashed) and together form the outer layer. The outer edges of the first and second layers, which overlap each other but do not overlap with the test substrate (shown as diagonal " / "), are joined together to form a cover or sleeve surrounding the test substrate. Figure 3 The side view depicted in the image shows how the inner test substrate is slightly smaller than the outer layer. Figure 3 Fasteners (e.g., Figure 3 The micro-hooks in the embodiments shown are positioned on the outer surface of the mask layer away from the subject's face and function to attach the mask insert to the mask worn by the subject. The overlapping ends of the first and second layers also engage with each other (in... Figure 1 and Figure 2This is referred to as a "pinch" in Chinese. The combination of the edges of the first and second layers also creates a barrier that blocks or significantly reduces airflow through the first and second layers and around the test substrate. Therefore, the combination of the first and second layers directs airflow toward the test substrate. The outer layer also acts as a protective layer for the test substrate and allows the mask insert to be handled and manipulated without direct contact with the test substrate. The outer layer includes materials for capturing bioaerosols smaller than 100 micrometers by providing a bioaerosol bandpass. The outer layer includes materials with very low pressure drop, allowing bioaerosols to flow easily through to the test substrate. The outer layer allows bioaerosols to pass through while preventing macroscopic contaminants (including ribonucleases and proteases) from contacting the inner test substrate, which could degrade analytes. The mask insert acts like a bandpass filter to allow larger bioaerosols to pass through the outer layer, while analytes are captured by the test substrate.

[0036] The mask insert has tiny hooks or adhesive strips along its edge, which allow the mask insert to attach to any face covering material and be easily removed without damaging the face covering.

[0037] The outer layer is perforated along the lines, so that when the user clamps both ends and pulls the outer layer outward, this portion of the outer layer is pulled apart and can be disposed of. This exposes the inner trapping substrate while allowing the user to still hold the outer substrate firmly at one end without contaminating the trapping substrate. The user can now place the inner substrate in a vial for testing.

[0038] The first and second layers can be made of the same or different materials. Suitable materials for making the first and second layers (which together form the outer layer) include, for example, spunbond polypropylene, spunbond polyester, meltblown fiber, carded nonwoven fiber, etc. Particularly suitable materials for making the first and second layers (which together form the outer layer) include 0.50 oz to 0.75 oz spunbond nonwoven cover material (available from Barry Global, Evansville, Indiana, USA). The materials of the first layer, the test substrate, and the second layer are designed to allow air exhaled by the subject to pass through the first layer, the test substrate, and the second layer. The material of the first layer, located near the subject's mouth and nose, is also designed to allow analytes contained in the air to pass through the first layer. The test substrate is designed to allow air to pass through (across) the test substrate, but also to trap analytes contained in the air exhaled by the subject. The second layer, located distal to the subject's face and proximal to the mask surface, also allows air to flow out from the mask insertion device. The appropriate fiber size for the material used to form the outer layer has an average fiber diameter ranging from about 5 micrometers to about 25 micrometers, including a range from about 10 micrometers to about 20 micrometers.

[0039] like Figures 1 to 3As shown, the mask insert preferably has a notch in the outer layer near one end of the mask insert, which allows for easier separation of a portion of the outer layer. Figure 1 The diagram also shows that the first and second layers include perforations that allow for easier separation of this portion of the outer layer from the test substrate. Preferably, a notch is formed in the outer substrate, its point ending at the starting point of the perforation. Figure 2 As further demonstrated, the separation of this portion of the outer layer exposes the test substrate. When both ends of the mask insert are grasped and pulled outward, this portion of the outer layer is pulled apart to expose the test substrate contained within the outer layer, while allowing the user to still hold one end of the mask insert through the remaining portion without contaminating the test substrate. The test substrate can then be processed to detect analytes captured on it. This portion of the mask insert's supply sleeve (in which the test substrate is housed) can be disposed of after the test substrate is removed. This allows for easy detachment of the outer substrate layer from the inner test substrate while the user still holds one end.

[0040] In another aspect, this disclosure relates to a mask insert that includes an integrated vertical flow measurement unit (VFA). Figure 4 An embodiment of a mask insert with an integrated vertical flow measurement unit is described. For example... Figure 4 As shown, the buffer well is positioned proximal to the notch and perforation to separate a portion of the outer layer, as described herein. This well allows the buffer solution added to the well to contact a portion of the test substrate. The buffer solution flows via capillary action toward a vertical flow measurement section located distal to the buffer well. In cases where a charged material (e.g., an electret) is used as the test substrate, the buffer solution short-circuits the charge of the electret material as it flows toward the VFA, thereby transporting the analyte to the VFA. The test substrate may be made of a hydrophobic material and / or a material less hydrophilic than the VFA pad, such that the flow is directed toward the VFA. The test substrate contacts the VFA to allow the transport of the buffer solution containing the analyte to the VFA. The buffer solution may also include other biomarkers that serve as positive and / or negative controls and can also be detected by the VFA. The VFA portion of the mask insert may include an observation “window” to allow observation of the VFA results.

[0041] Figure 5 An embodiment of a mask insert with an integrated vertical flow measuring unit is described, the mask insert having a positioning on Figure 5 The buffer dispensing device is depicted near the notch and perforation. The buffer dispensing device provides buffer orifices (in...) Figure 4This is an alternative or supplement to (as depicted in the image). The buffer dispensing device may be an ampoule embedded within the mask insert. When pressure is applied to the mask insert where the buffer dispensing device is positioned, all or part of the buffer dispensing device ruptures to release the buffer contained within. The buffer flows toward a vertical flow measurement section located distal to the buffer dispensing device via capillary action. In the case where a charged material (e.g., an electret) is used as the test substrate, the buffer short-circuit the charge of the electret material as it flows toward the VFA, thereby transporting the analyte to the VFA. The test substrate may be made of a hydrophobic material and / or a material less hydrophilic than the VFA pad, such that the flow is directed toward the VFA. The test substrate contacts the VFA to allow the transport of the buffer containing the analyte to the VFA. The buffer may also include other biomarkers that serve as positive and / or negative controls and may also be detected by the VFA. The VFA portion of the mask insert may include an observation “window” to allow observation of the VFA results.

[0042] Figure 6 Depicting Figure 5 Another embodiment of the mask insert shown in the figure, but the mask insert has a removable outer layer. Figure 6 A VFA for single analyte detection and a VFA for performing multiple tests are described. Removing the outer layer of the mask insert between the buffer dispensing device and the VFA allows buffer to flow through the test substrate without also flowing into the material forming the outer layer.

[0043] In another embodiment, the mask insert may include two or more buffer dispensing devices. Figure 7 An exemplary embodiment of a mask insert is depicted, the mask insert including three buffer dispensing devices. Figure 7The embodiments shown also include a removable outer layer. Buffer from the buffer dispensing device flows to the VFA, but barriers and / or closed, non-porous spacers (or dividers) in the test substrate allow independent elution and extraction of the analyte from the test substrate “channels” and prevent cross-flow of each buffer into other “channels” of the test substrate intended to detect different analytes. The buffer flows towards a vertical flow measurement section located distal to the buffer dispensing device via capillary action. When a charged material (e.g., an electret) is used as the test substrate, the buffer short-circuits the charge of the electret material as it flows towards the VFA, thereby transporting the analyte to the VFA. The test substrate may be made of a hydrophobic material and / or a material less hydrophilic than the VFA pad, such that the flow is directed towards the VFA. Removing the outer layer of the mask insert between the buffer dispensing device and the VFA allows the buffer to flow through the test substrate without also flowing into the material forming the outer layer. The test substrate contacts the VFA to allow the transport of the buffer containing the analyte to the VFA. The buffer may also include other biomarkers that serve as positive and / or negative controls and can also be detected by the VFA. The VFA portion of the mask insert may include an observation “window” to allow observation of the VFA results. Figure 7 Exemplary embodiments of multiple tests are also described.

[0044] Figure 8 Another exemplary embodiment of a mask insert is depicted, which includes three buffer dispensing devices. Figure 8 The embodiments described also include perforations proximal to these buffer dispensing devices and proximal to the VFA to allow removal of only the outer layer surrounding the central portion of the test substrate. Buffer from these buffer dispensing devices flows toward the VFA, but barriers and / or spacers (or partitions) in the test substrate prevent each of the buffers from cross-flowing into the test substrate. The buffer flows toward a vertical flow measurement section located distal to the buffer dispensing device via capillary action. When a charged material (e.g., an electret) is used as the test substrate, the buffer short-circuits the charge of the electret material as it flows toward the VFA, thereby transporting the analyte to the VFA. Removing the outer layer of the mask insert between the buffer dispensing device and the VFA allows the buffer to flow through the test substrate without also flowing into the material forming the outer layer. The test substrate may be made of a hydrophobic material and / or a material less hydrophilic than the VFA pad, such that the flow is toward the VFA. The test substrate contacts the VFA to allow transport of the buffer containing the analyte to the VFA. The buffer may also include other biomarkers used as positive and / or negative controls and may also be detected by the VFA. The VFA portion of the mask insert may include an observation “window” to allow observation of the VFA results. Figure 8 Exemplary embodiments of multiple tests are also described.

[0045] In another embodiment, the mask insert includes an integrated vertical flow measurement unit and at least one integrated delivery plunger for introducing buffer solution into the test substrate. Figure 9A An exemplary embodiment with three integrated delivery plungers is depicted. Figure 9B The delivery plunger is depicted. The vertical rod on the right is a locking mechanism that can be flipped upwards to allow the plunger on the left to be depressed, thereby discharging the buffer solution from the plunger cavity into the test substrate.

[0046] In another embodiment, the mask insert includes an integrated sideflow measuring unit. Figure 10 An exemplary embodiment of a mask insert having an integrated sideflow measurement unit (LFA) is depicted. Figure 10 Examples include pores for introducing buffer solution to extract and / or elute analytes and transport analytes from the test substrate toward the sample pad of the side-flow measurement section. For example... Figure 10 As depicted, the test substrate contacts the sample pad of the LFA. The LFA can be in the form of a standard LFA box or integrated as a laminated LFA with a mask insert. Figure 10 The embodiments described herein can also be used for multiple tests.

[0047] exist Figure 11 In another embodiment depicted herein, the mask insert includes an integrated lateral flow measurement unit and further includes a buffer dispensing device (ampoule) for introducing buffer solution to extract and / or elute analytes and transport the analytes from the test substrate toward the sample pad of the lateral flow measurement unit. As discussed herein, pressure applied to the buffer dispensing device releases the buffer solution, which flows through the test substrate toward the LFA via capillary action. Figure 11 As depicted, the test substrate contacts the sample pad of the LFA. The LFA can be in the form of a standard LFA box or integrated as a laminated LFA with a mask insert. Figure 11 The embodiments described herein can also be used for multiple tests.

[0048] exist Figure 12 In another exemplary embodiment depicted herein, the mask insert includes an integrated lateral flow measurement unit and further includes a buffer dispensing device (ampoule) for introducing buffer solution to extract and / or elute analytes and transport the analytes from the test substrate toward the sample pad of the lateral flow measurement unit. As discussed herein, pressure applied to the buffer dispensing device releases the buffer solution, which flows through the test substrate toward the LFA via capillary action. Removing the outer layer of the mask insert between the buffer dispensing device and the LFA allows the buffer solution to flow through the test substrate without also flowing into the material forming the outer layer. Figure 12As depicted, the test substrate contacts the sample pad of the LFA. The LFA can be in the form of a standard LFA box or integrated as a laminated LFA with a mask insert. Figure 12 The embodiments described herein can also be used for multiple tests.

[0049] The test substrate is designed to capture analytes from air samples taken from a subject. In some embodiments, the test substrate has a pore size smaller than the analyte size. In other embodiments, the test substrate is made of a material that attracts analytes. In some embodiments, the test substrate has a pore size smaller than the analyte size and is also made of a material that attracts analytes. The pore size used to capture the analyte is less than 100 micrometers. Particularly suitable pore sizes for capturing analytes (e.g., pathogens) are less than 75 micrometers, less than 50 micrometers, and less than 25 micrometers. The material can be changed to target a specific analyte while minimizing pressure drop. For example, capturing COVID may require a higher pressure drop because COVID is smaller and therefore requires a denser material. TB can be captured with a less dense material at similar efficiency, thus having a lower pressure drop and higher throughput. Exemplary analytes intended to be captured by the test substrate of a mask insert include COVID-19 (approximately 100 nm), influenza (approximately 80 nm to 120 nm); mycobacteria (approximately 7 micrometers); and other bioaerosols containing respiratory pathogens (less than 10 micrometers). As described herein, test substrate materials can be treated (e.g., by imparting a charge to the material) to “attract” the analyte to the test substrate. Thus, for example, less dense materials can be treated to attract COVID to the test substrate compared to untreated materials.

[0050] A mask insert is attached (attached) to the inner surface of the mask. The mask insert is attached to the mask surface by adhesive and other fasteners (e.g., hook-and-loop fasteners). Preferably, the mask insert includes micro-hooks that allow it to attach to the inner surface of the mask worn by the subject. Other attachment mechanisms are also suitable. For example, adhesives can be used to attach the mask insert to the mask surface. Generally, non-permanent adhesives are preferred, allowing the mask insert to be removed from the mask surface.

[0051] Subjects expel air through breathing, coughing, sneezing, talking, and combinations thereof.

[0052] In one embodiment, the mask insert is reversibly attached to a mask worn by a user. As used herein, “reversibly attached” means the ability to attach the mask insert to the mask, then remove the mask insert from the mask, and then reattach the mask insert to the mask. In some embodiments, the mask is reusable, and a new mask insert is attached to the mask.

[0053] In another embodiment, the mask insert is semi-permanently attached to the mask worn by the user. In this embodiment, the mask insert may be detached from the mask after being attached to the mask surface, but is not intended to be reattached to the mask insert after being removed from the mask surface.

[0054] In another embodiment, the mask insert is permanently attached to the mask worn by the user. In this embodiment, once the mask insert is attached to the mask surface, the mask insert is not intended to be detached from the mask.

[0055] Generally, a mask insert (whether reversibly or permanently attached) is positioned on the inner surface of the mask, through which airflow is caused by the user exhaling air via normal breathing, forceful breathing, coughing, sneezing, or any combination thereof, towards the mask insert.

[0056] As used herein, a test substrate (also referred to herein interchangeably as a "test substrate" and a "collection substrate") may include any inanimate surface that can come into contact with an individual or a group of individuals, and that inanimate surface is capable of containing pathogens on its outer surface. For example, in one embodiment, the test substrate may be an intact, complete face mask or a face mask from which a portion of the face mask (i.e., the test substrate) has been removed. The substrate may also include a removable face mask insert. In some embodiments, the face mask insert may be cleaned and reused by being placed back into the face mask.

[0057] In some embodiments, the test substrate is treated to extract and / or remove pathogen material from the test substrate. In other embodiments, the test does not require the removal or extraction of pathogen material from the test substrate. For example, the test substrate can be analyzed by placing it in a reaction solution (e.g., buffer and / or water) that results in a colorimetric reaction indicating the presence or absence of a pathogen. In another embodiment, the test substrate is placed in a reaction solution (e.g., buffer and / or water), thereby dissolving the test substrate. In another embodiment, a reagent is applied to the test substrate. In yet another embodiment, the reagent is brought into contact with the test substrate, which draws the reagent into the test substrate via capillary action.

[0058] The test substrate can be suitably made of synthetic fibers, natural fibers, and combinations thereof. The fibers used to form the layer can be hydrophobic fibers, hydrophilic fibers, and combinations thereof. Hydrophobic fibers include, for example, polylactone, poly(caprolactone), poly(L-lactic acid), poly(glycolic acid), similar copolymers (alkyl acrylates), polybutadiene, polyethylene, polystyrene, polyacrylonitrile, polyethylene glycol (terephthalate), polysulfone, polycarbonate, poly(vinyl chloride), and combinations thereof. Hydrophilic fibers include, for example, linear poly(ethyleneimine), cellulose, cellulose acetate and other grafted cellulose, poly(hydroxyethyl methacrylate), poly(ethylene oxide), polyvinylpyrrolidone, poly(acrylic acid), poly(ethylene glycol), poly(vinyl alcohol), poly(vinyl acetate), poly(acrylamide), proteins, poly(vinylpyrrolidone), poly(styrene sulfonate), and combinations thereof. Other suitable fibrous materials include, for example, acrylonitrile / butadiene copolymer, cellulose, cellulose acetate, chitosan, collagen, DNA, fibroin, fibronectin, nylon, poly(acrylic acid), poly(chlorostyrene), poly(dimethylsiloxane), poly(etherimide), poly(ether sulfone), poly(ethyl acrylate), poly(ethyl vinyl acetate), poly(ethyl-co-vinyl acetate), poly(ethylene oxide), poly(ethylene terephthalate), poly(lactic-co-glycolic acid), poly(methacrylate), poly(methyl methacrylate), poly(methylstyrene), poly(styrene sulfonate), poly(styrene sulfonyl fluoride), poly(methyl methacrylate ... (Styrene-co-acrylonitrile), poly(styrene-co-butadiene), poly(styrene-co-divinylbenzene), poly(vinyl acetate), poly(vinyl alcohol), poly(vinyl chloride), poly(vinylidene fluoride), polyacrylamide, polyacrylonitrile, polyamic acid (PAA), polyamide, polyaniline, polybenzimidazole, polycaprolactone, polycarbonate, polydimethylsiloxane-co-ethylene oxide, polyether ether ketone, polyethylene, polyethyleneimine, polyimide, polyisoprene, polylactide, polypropylene, polystyrene, polysulfone, polyurethane, polyvinylpyrrolidone, protein, SEBS copolymer, filament, styrene / isoprene copolymer and combinations thereof. 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), poly(hydroxystyrene) blend-poly(ethylene oxide), and combinations thereof.

[0059] Another suitable layer for the test substrate can be an electret (including thermal electrets and fibrillated electret films). An electret is a dielectric material with quasi-permanent charge or dipole polarization. Electrets are commercially available. Electrets can be prepared by heating the material while simultaneously exposing it to an electric field, thereby orienting many dipoles in the material in a preferred direction. After heating, the material is "frozen" and can maintain the position of its electric dipoles for a long time. Suitable materials for preparing electrets include, for example, materials that can now be used to manufacture thermal electrets, including organic materials (e.g., hard rubber, naphthalene, polymethyl methacrylate, and many polymers) and inorganic materials (e.g., sulfur, quartz, glass, talc, and some ceramics). Electret fiber films are particularly suitable. Polyvinylidene fluoride (PVDF) / polytetrafluoroethylene (PTFE) NP electret nanofiber films can be formed by electrospinning. Fibrillated electret films, as described in Van Turnhout (US Patent 3,998,916), are also suitable.

[0060] A particularly suitable test substrate is a charged material (e.g., polypropylene, polylactic acid, and electret) that attracts analytes contained in an air sample. Therefore, due to the size difference between the test substrate and the analyte, the analyte contained in the subject's air is not necessarily captured by the test substrate, but rather attracted to it.

[0061] Examples of using polypropylene as a test substrate advantageously provide an inert substrate that allows biological materials (e.g., pathogens) to remain stable over extended periods, including when mask inserts and / or test substrates need to be transported for testing. The test substrate may also include coatings and chemical treatments that allow for easier elution of analytes. Materials with lower surface energy can be used to increase the wettability of the test substrate material, thus requiring less buffer solution to wet the material and consequently concentrating the analyte in the elution buffer. The advantage of hydrophobic polypropylene is that it can be easily expelled from nonwoven fabrics and allows for the elution of viruses by simply applying a buffer solution (e.g., 0.1% Triton X-100) that performs coarse extraction against certain pathogens (e.g., SARS-CoV-2).

[0062] It can also process test substrate materials to impart charges, make the materials more or less hydrophilic, make the materials more or less hydrophobic, and combinations thereof.

[0063] Suitable test substrate materials also include materials that are soluble and / or liquid-soluble. For example, cellulose acetate nanofibers are capable of dissolving upon contact with a liquid. It should be understood that the entire test substrate and / or portions thereof (e.g., collection substrate) can be soluble or liquid-soluble. Advantageously, the substrate itself can be completely dissolved, thereby releasing all analyte material into the eluent without the need for a removal process. Suitablely, the substrate can be prepared during substrate dissolution, substrate reaction with and stabilization of RNA, or the performance of some other service assay. Suitablely, the substrate can be inert before it dissolves. Additionally, or alternatively, the protective inert layer of the dissolved inner substrate exposes a less inert, active inner substrate with active diagnostic properties. For example, the test substrate (e.g., a rod) is placed in an eluent that dissolves the outer coating, thereby releasing the full amount of viral material. The test reagent then dissolves and interacts with the analyte material.

[0064] Fibers can be subnanofibers, nanofibers, microfibers, and combinations thereof, with diameters ranging from subnanometers to micrometers. Subnanofibers can be prepared using template-assisted growth, directed attachment growth, ligand-controlled growth, and catalyst-guided growth. Subnanofibers (also referred to herein as “subnanofibers”) refer to one-dimensional structures with ultra-high aspect ratios and diameters ranging from a few angstroms to tens of angstroms. Ultrafine nanofibers with diameters less than 100 nm can be produced by phase separation, self-assembly, island-of-situ methods, template synthesis, electrospinning, and bubble electrospinning. Nanofiber diameters range, for example, from about 3 nm to less than 1 μm. Microfiber diameters range, for example, from 1 μm to about 10 μm. Nanofibers and microfibers can be produced using phase separation, self-assembly, island-of-situ methods, template synthesis, electrospinning and bubble electrospinning, meltblowing, spin-blowing, and other methods. Fiber diameters and morphologies can be determined using scanning electron microscopy (SEM), transmission electron microscopy (TEM), and atomic force microscopy (AFM). The test substrate fibers can be woven, knitted, crocheted, knotted, pressed together, interwoven, combined, bound together in the form of sheet webs, and combinations thereof. Spunbond materials prepared with continuous fibers can be used to form layers, which are formed by continuously extruding polymers through a spinneret to form discrete filaments. The filaments can then be mechanically or pneumatically stretched without breaking to orient the polymer filaments. The continuous filaments can be deposited onto a carrier belt in a substantially random manner to form a web. Meltblown fiber forming layers can also be used. Electrospun fiber forming layers can also be used. As known to those skilled in the art, electrospinning involves typically stretching individual polymer chains in a polymer solution into nanoscale or submicron-scale structures in fibrous form through nozzles / orifices with very small diameters at high voltage.

[0065] The test substrate layer may include additives. Suitable additives include, for example, antimicrobial additives (e.g., silver-containing antimicrobials and antimicrobial peptides), analgesic compounds (e.g., lidocaine), antibiotics (e.g., neomycin), thrombotic compounds, nitric oxide-releasing compounds (e.g., stenoimide and NO-complexes), bactericidal compounds, fungicidal compounds, antimicrobial compounds, other pharmaceutical compounds, adhesives, fragrances, odor-absorbing compounds, preservatives, ribonuclease inhibitors, protease inhibitors, and nucleic acids (including deoxyribonucleic acid, ribonucleic acid, and nucleotide analogs).

[0066] Suitable analytes are contained in gases and aerosol droplets in the air exhaled by the user. Suitable analytes include microorganisms, chemicals, proteins, nucleic acids, and combinations thereof. Suitable microorganisms include bacteria and viruses.

[0067] Particularly suitable microorganisms include pathogens. The term "pathogen" is used in its general sense to refer to bacteria, viruses, and other microorganisms that cause disease directly or indirectly. Exemplary pathogens include, for example, Yersinia spp., Klebsiella spp., Providenella spp., Erwinia spp., Enterobacter spp., Salmonella spp., Serratia spp., Aerobacter spp., Escherichia spp., Pseudomonas spp., Shigella spp., Vibrio spp., Aeromonas spp., Streptococcus spp., Staphylococcus spp., Micrococcus spp., Moraxella spp., Bacillus spp., Clostridium spp., Corynebacterium spp., Ebola spp., Francisella spp., Haemophilus spp., Bacteroides spp., Listeria spp., and Erysipelothrix rhusiopathiae. Genus, Acinetobacter, Brucella, Pasteurella, Flavobacterium, Fusobacterium, Streptococcus, Coleobacterium, Legionella, Treponema, Leptospira, Actinomyces, Nocardia, Rickettsia, Micrococcus, Mycobacterium, Neisseria, Campylobacter, Pathogenic viruses (e.g., papillomavirus, parvovirus, adenovirus, herpesvirus, vaccine virus, arenavirus, coronavirus, rhinovirus, respiratory syncytial virus, epidemic...) Common cold virus, parvovirus, paramyxovirus, reovirus, retrovirus, rhabdovirus, human immunodeficiency virus (HIV), tapeworm, hymenolepis, sparganum, echinococcosis, fascioliasis, heterocerca, clonorchis, fascioliasis, schistosomiasis, pinworm, whipworm, roundworm, hookworm, nematode, brucea, loa worm, onchocerca, dragon worm, Naegleria, Acanthamoeba, malaria parasite, trypanosoma, leishmaniasis Shimania, Toxoplasma gondii, endoamyotrophic lateral sclerosis, Giardia lamblia, Isosporidia, Cryptosporidium, Enterocytozoa, Strongyloides, Trichinella, fungi (which cause, for example, tinea, histoplasmosis, blastomycosis, aspergillosis, cryptococcosis, sporotrichosis, coccidioidomycosis, paracoccidioidomycosis, mucormycosis, candidiasis, dermatomycosis, prosomal diseases, pityriasis, mycoidosis, paracoccidioidomycosis, daturiasis, pseudoalishiasis, trichosporidiosis, pneumosporidiosis) and combinations thereof.

[0068] Particularly suitable chemicals include ketones, nicotine, cocaine, opioids, cannabis, benzodiazepines, amphetamines, barbiturates, and combinations thereof.

[0069] It can also detect proteins, DNA, and RNA.

[0070] Suitable face masks include any face covering worn over or in front of a user's mouth and nasal passages, where air is expelled from the user, for example, during exhalation, when speaking, coughing, and sneezing. Face masks can be secured to the user's face using straps, bands, straps, and combinations thereof. Face masks also include face shields. Face masks also include headscarves, neck warmers, scarves, towels, and fabric coverings positioned over the user's nose and mouth. Therefore, the face mask inserts disclosed herein can be coupled to any type of face covering such that at least a portion of the exhaled air (through breathing, coughing, sneezing, speaking) is delivered to a device in which analytes contained in an air sample can be collected by a test substrate.

[0071] When the analyte is a pathogen, the amount of pathogen particles captured can be expressed as filtration efficiency, minimum efficiency reporting value (MRC) level, and microparticle performance level. Appropriately, the filtration efficiency of the test substrate ranges from about 60% to about 95%. Appropriately, the MRC level is at least 12. Appropriately, the microparticle performance level is at least 1900.

[0072] Any suitable method can be used to clean the face mask and / or face mask insert, such as by heat treatment, ultraviolet light irradiation, and other disinfection methods. In other embodiments, the face mask and / or face mask insert is a disposable face mask or face mask insert. In these embodiments, the face mask and / or face mask insert is destroyed after removal.

[0073] In one aspect, this disclosure relates to a face shield for testing pathogen infection. The face shield includes a test substrate that includes a trapping agent for capturing pathogens.

[0074] Suitable capture agents include, for example, antibodies that specifically bind to pathogens, and ligands that specifically bind to pathogens (e.g., surface molecules that pathogens must bind to in order to infect their hosts, such as sugars, glycoproteins, etc.). Capture agents can be covalently or non-covalently linked to the test substrate via linkers. 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.

[0075] In one embodiment, a mask worn by an individual covers the subject's mouth and nose, such that when the individual breathes, coughs, and / or sneezes into the mask, pathogens are trapped and / or adsorbed by the mask material. The mask may include ear loops and / or straps to secure it to the individual wearer's face. The mask may include strips of material attached to and extending along each side of the mask to provide an enhanced fluid seal between the mask's periphery and the wearer's face.

[0076] The test substrate can be coated and / or treated to absorb and / or retain pathogens.

[0077] The face mask and / or the mask insert portion of the face mask can be configured to maximize the surface area of ​​the test substrate. For example, the test substrate may include particles, such as microparticles, nanoparticles, and beads. The particles may be coated with a trapping agent.

[0078] Suitable, the test substrate portion of the mask and / or mask insert may be an immunochromatographic test substrate, a colloidal gold test substrate, or a combination thereof. The substrate may include, for example, quantum dot-labeled test substrates, colloidal gold-labeled test substrates, colloidal selenium-labeled test substrates, upconversion phosphorescent-labeled test substrates, nano-rare earth fluorescent complex-labeled test substrates, time-resolution chromatography test substrates, chemiluminescence test substrates, and other test substrates.

[0079] In embodiments where the face shield is intended to include a test substrate, the face shield is configured to receive the test substrate. For example, the face shield may be perforated, and the test substrate may be inserted into the face shield at the location of the perforation. In other embodiments, the test substrate may be attached to the face shield, for example, by hook-and-loop fasteners and adhesives.

[0080] The mask insert may also include a tab for marking the mask insert and handling the mask without contaminating the test substrate.

[0081] This can form a mask insert, and particularly a perforated portion of the test substrate, allowing the test substrate to be divided into different parts (or tabs), such as... Figure 1 As shown in the diagram. Perforations can be present between each tab. Additionally, or alternatively, the perforations can be parallel to the top (or bottom) edge of the test substrate. Figure 1 A and Figure 1 As shown in B, in addition to the perforations between each tab, the perforations can be oriented such that individual tabs can be separated from the label portion, allowing individual tabs to separate independently of the other tabs on the test substrate, thus keeping the remaining tabs attached. Additionally, or alternatively, the test substrate can be perforated along its top (or bottom) edge, and the space (or gap) can separate each tab, allowing each tab to be independently attached to the top or bottom edge of the test substrate, as shown in Figure B. Figure 1 As shown in B. In Figure 1 In another embodiment shown in C, the tabs can "float freely" within the test substrate. For example, the test substrate can have four separate tabs that are not connected but are contained in the same plane of the test substrate, instead of a single strip with perforations that produce the four tabs. Figure 1 A to Figure 1The exemplary embodiment shown in C illustrates four patch portions that can be tested individually. For example, patch 1 can be assembled with other patches obtained from multiple patients. Patch 2 can then be tested individually. Patch 3 can be used as a control. Patch 4 can be stored for future testing. Although Figure 1 An exemplary embodiment with four tabs is shown, but it should be understood that the test substrate may have fewer than four tabs and more than four tabs. Figure 2 The user's face mask was shown, which had a test substrate with four test tabs.

[0082] It should be understood that "patch" can refer to a single acquisition substrate as well as a separable portion of a test substrate. For example, a test substrate patch may include one or more protective layers, with the acquisition substrate sandwiched between the protective layers (and other layers). In this embodiment, the test substrate patch (i.e., a portion of the test substrate) can be separated from the remaining test substrate, thus undisturbed by other patch portions. In another embodiment, the acquisition substrate patch may be sandwiched between the protective layers. In this embodiment, the acquisition substrate patch can be removed independently from between the protective layers without the protective layers(s) and / or without contacting or interfering with the remaining acquisition substrate patch. The advantage of the "teeth" embodiment is that the clinician can simply hold the patch, tear along the top perforation, thereby releasing both the other layers and the inner layer from the common spine, and the inner layer can be simply stored in a container for further testing. Nothing is contaminated, the outer layer is disposed of, etc. Patches may also include multiple layers, as described herein with respect to multi-layer test substrates. Patches(s) may share a common spine but are separate (e.g., as described herein with respect to multi-layer test substrates). Figure 1 (As shown in C). The tabs can be perforated for easy removal without contamination.

[0083] The test substrate can also be a multilayer test substrate. A multilayer test substrate has an inner collection substrate that is protected on all sides by additional layers. The inner collection layer is made of a material with a high surface area for collecting the pathogen to be detected. The inner collection substrate can also be released from the protective layers. These protective layers and the inner collection substrate can be made of different materials designed for the specific purpose of the layer (e.g., protection and sample collection). For example, the inner collection substrate can be a corrugated, double-sided polyester swab material. The inner collection substrate can be coated with reagents to maintain pathogen load collection. The inner collection substrate can also be coated with reagents to stabilize pathogen material. Figure 3A multi-layer test substrate is depicted. The protective layer can also function as a transport layer, which is porous and flexible, enabling it to receive and temporarily store exhaled microdroplets of liquid before being transferred and absorbed by the underlying collection layer(s). The protective layer can be a breathable, touch-protective coating. The protective layer positioned furthest from the user's face blocks all airflow, while the protective layer positioned closest to the user's face allows airflow.

[0084] In some embodiments, the inner acquisition substrate(s) can be separated from the protective layers. In some embodiments, the protective layers can be opened to release or expose the inner acquisition substrate. For example, in an exemplary embodiment having a proximal protective layer and a distal protective layer and the inner acquisition substrate positioned between these protective layers, the protective layers may be larger than the inner acquisition substrate. When clamped together, the surfaces of these protective layers can contact each other and be held together using adhesives, press-fit locking zippers, tape, or hinges. Separation of the protective layers allows access to the inner acquisition layer, which can then be removed for testing. Separation of the protective layers also allows for the insertion or replacement of unused inner acquisition substrates.

[0085] Features of multilayer test substrates and multilayer tabs include, for example, (multiple) outer layers protecting inner layers from contamination, (multiple) inner layers being double-sided, (multiple) inner layers including adhesives, (multiple) inner layers including stabilizers, (multiple) inner layers including test layers, and (multiple) inner layers being multilayered.

[0086] In one embodiment, a multilayer test substrate may include one or more reagent layers (also referred to herein as “reagent material” comprising analytical reagents). As used herein, “analytical reagent” refers to a component used to detect pathogens. For example, analytical reagents may include, for example, buffer components, salts, dNTPs, oligonucleotide primers, polymerases, reverse transcriptases, and combinations thereof. Analytical reagents may suitably be lyophilized, in liquid form, in gel form, or combinations thereof. The reagent layer comprising the analytical reagent may be separated from the other layers of the test substrate by a coating to prevent the analytical reagent from contacting the collection layer and / or becoming activated until such a time when the test substrate will be processed for analysis. The analytical reagent may be activated, for example, by placing the test substrate (and / or the collection layer having a soluble layer comprising the analytical reagent) in a liquid medium (e.g., a buffer comprising water), thereby dissolving the coating to release the analytical reagent, which may then also dissolve in the buffer. The coating may be fusible, thereby allowing temperature adjustment such that the coating melts to release the analytical reagent and can form a mixture by which pathogens can be detected. The reagent layer may include microparticles and / or beads containing the analytical reagent.

[0087] In some embodiments, the test substrate and(s) of the internal sampling substrate(s) may be packaged in sterilized packaging to prevent contamination during storage and handling. The packaging may be opened to allow the test substrate or internal sampling substrate to then be placed in a face shield and / or together with a protective layer.

[0088] Multilayer test substrates may include spacers between individual layers. These spacers may be positioned at the edges of the sampling area to prevent airflow through the spacer material. Additionally or alternatively, the spacers may be materials through which air and pathogen particles can pass.

[0089] Multilayer test substrates may include labels (see example) Figure 1 The label can be a blank area on which information can be printed. The label can also be pre-printed or have a machine-readable barcode. The information included on the label can be used to identify the test substrate (including individual test substrate patches), to track the collected test substrate, and to associate it with the user.

[0090] In a multilayer test substrate, individual layers can have pore sizes different from other layers. In one exemplary embodiment, the first protective layer can have pore sizes with low filtration efficiency, allowing pathogen-sized particles to pass freely through the protective layer to reach the collection layer.

[0091] In an exemplary embodiment, the test substrate may have: a first protective layer made of a fibrous material having a pore size range that allows pathogen-sized particles to pass freely through the first protective layer; and at least one collection layer made of a different fibrous material having a pore size that allows particles smaller than the pathogen size to be captured by the collection layer. Suitablely, the collection layer itself may be a multilayer laminate to increase the viral load captured per test. If the inner substrate patch can capture a viral density of X, then there should exist a function f(Ln) = Ln x viral load per layer, where Ln is the number of layers. It should be understood that efficiency decreases by adding layers.

[0092] Multiple collection layers can be treated to enhance the capture of pathogen particles. In one embodiment, the collection layer is treated with an adhesive to which pathogen particles are adhered. In another embodiment, the collection material is treated with a capture ligand. In yet another exemplary embodiment, the collection material is treated with a combination of an adhesive and a capture ligand. The capture ligand may be an antibody (including antibody fragments), an aptamer, a magnetic particle, or other ligand types that can bind to a pathogen of interest. The capture ligand is intended to attract and retain the pathogen, enabling subsequent analyses to be performed to detect and / or identify the pathogen.

[0093] An exemplary multilayer test substrate includes, for example, a collection layer without a trapping agent and without a protective layer. Another exemplary multilayer test substrate includes a protective layer and a collection layer. Yet another exemplary multilayer test substrate includes a protective layer and a collection layer, wherein the collection layer includes a trapping agent (e.g., an adhesive, a trapping ligand, or a combination thereof). Yet another exemplary multilayer test substrate includes a protective layer and multiple collection layers.

[0094] The protective layer can be a material that allows airflow and protects the inner acquisition layer. The protective layer can be treated or coated with reagents such as antimicrobial agents, ribonuclease inhibitors, protease inhibitors, preservatives, and combinations thereof. For example, the protective layer also provides physical protection to the inner acquisition substrate from contact. Each layer can be perforated to allow for the separation of the splice.

[0095] The orientation of the test substrate can be flat (e.g., in paper sheets), coiled, rod-shaped, hollow cylindrical, honeycomb, or combinations thereof. Coiled, rod-shaped, and hollow cylindrical substrates can be axial or perpendicular to the breathing direction. Coiled, rod-shaped, and hollow cylindrical substrates can also be easily slid into the tube for storage and / or handling. It should be understood that each of these embodiments provides a three-dimensional structure to enhance acquisition and allow airflow.

[0096] In one aspect, this disclosure relates to a method for detecting the presence of a pathogen. In one embodiment, the method detects the presence of a pathogen on a non-living surface that may come into contact with an individual or a group of individuals. In another embodiment, the method detects the presence of a pathogen infection in a group of individuals, wherein at least one individual in the group has or is suspected of having a pathogen infection. The method includes: collecting test substrates worn by each individual in the group of individuals; combining the test substrates collected from each individual to form a pooled sample of test substrates; and performing pathogen analysis on the pooled sample of test substrates, wherein detecting the presence of a pathogen in the pooled sample of test substrates indicates that at least one individual in the group of individuals has been in close contact with the pathogen, is infected with the pathogen, or may require further diagnostic testing.

[0097] The method may further include performing a second test independently on each individual in the population to identify individuals infected with the pathogen.

[0098] In some embodiments, the method is repeated for subgroups or subpools of a first group of individuals. By way of example only, the method of this embodiment includes: collecting test substrates worn by each individual in the first group of individuals; combining the test substrates collected from each individual to form a first pooled sample of test substrates; performing pathogen analysis on the first pooled sample of test substrates, wherein the detection of the presence of a pathogen in the pooled sample of test substrates indicates that at least one individual in the group of individuals has been in close contact with the pathogen, is infected with the pathogen, or may require further diagnostic testing; collecting test substrates worn by each individual in a second group of individuals, wherein the second group of individuals is a subgroup of the first group of individuals; combining the test substrates collected from each individual to form a second pooled sample of test substrates; performing pathogen analysis on the second pooled sample of test substrates, wherein the detection of the presence of a pathogen in the pooled sample of test substrates indicates that at least one individual in the group of individuals has been in close contact with the pathogen, is infected with the pathogen, or may require further diagnostic testing. This method can be repeated any number of times to identify individuals or subgroups of individuals infected with the pathogen, or material substrates that have been in contact with individuals or subgroups of individuals infected with the pathogen.

[0099] This document describes suitable test substrates. Preferably, the test substrate is a face mask or a portion thereof (e.g., a face mask insert) as described herein.

[0100] This article describes suitable pathogens to be tested.

[0101] In one aspect, this disclosure relates to a method for detecting the presence of a pathogen infection in an individual who has or is suspected of having a pathogen infection, the method comprising: collecting a test substrate worn by the individual from the individual; and performing pathogen analysis on the test substrate, wherein detecting the presence of a pathogen indicates that the individual has been in close contact with the pathogen, is infected with the pathogen, or may require further diagnostic testing.

[0102] This document describes suitable test substrates. Preferably, the test substrate is a face mask or part of a face mask as described herein (e.g., face mask insert, test substrate, patch).

[0103] In some embodiments, the pathogen(s) and / or pathogen material to be tested are extracted (eluted or removed) from the test substrate. In some embodiments, the test substrate can be analyzed directly, including, for example, a soluble test substrate or an evaporable test substrate used (including a patch, an inner test substrate layer, etc.).

[0104] This article describes suitable pathogens to be tested.

[0105] The test substrate used in the methods disclosed herein can be tested directly or removed from a mask (e.g., by cutting the mask to remove a portion of the mask to be tested), or the test substrate in the form of a mask insert can be configured for easy use with an analyzer. Any analyzer used to test the substrate for the presence of an analyte (e.g., the pathogen in this disclosure) is suitable for use in this disclosure. In one embodiment, the test substrate can be inserted into the analyzer, and the pathogen can be detected by the analyzer. In other embodiments, it should be understood that the analyzer can be a point-of-care analyzer, in which the detection reagent comes into direct contact with the test substrate, such as by dropping a solution comprising the detection reagent directly onto the test substrate. These point-of-care analyzers can be used by individuals at home, at the workplace, etc. The signal output of the analyzer can provide information about the presence of pathogens in the sample. For example, the signal output from the analyzer's detector can be transmitted to a data processor for storage, processing, and analysis. Appropriately, the test substrate (including a patch) can be placed in an ion mobility spectrometer and the pathogen can be tested.

[0106] like Figure 4 As shown, the test substrate is compatible with additional sample preparation methods, such as mechanical, chemical, and force application (e.g., magnetic force).

[0107] Figure 5 A flowchart illustrating sample detection using amplification is shown. After collection, RNA is extracted from the sample and subjected to reverse transcription to produce a cDNA pool. The cDNA is then amplified using primers specifically targeting the pathogen of interest, employing, for example, polymerase chain reaction (“PCR”). Following amplification, the PCR products can be further analyzed. Analysis may include simple product / no-product results and / or sequencing of the PCR products. The amount of product can also be used to determine the concentration of the pathogen.

[0108] After sample collection via a mask and / or test substrate, the collected samples can be analyzed using commercially available equipment and methods.

[0109] The range of virus density and viral load capacity in the above device embodiments can be determined by utilizing the ratio of controlled spiked tests based on the inside of the mask.

[0110] The approximate range of viral loads collected by embodiments of the test substrate device can be inferred using the following assumptions, assuming that an infected person wears the device under different conditions: VD d1 / VD m1 :VD d2 / VD m2 Where VD = based on millimeters per square millimeter (mm) 2The normalized viral density of the inner substrate is used to determine the viral load; d1 = pathogen-spiking device in the mask; m1 = pathogen-spiked mask with the device; d2 = inferred performance of the device embodiment; m2 = mask collected from a detached patient. Once the range of results from the pathogen-spiking test and the collected masks is determined, an approximate range of the collection potential of the device embodiment can be inferred. In another example, the test substrate device can be doped with a range of viral loads to simulate low, average, and high densities, thereby inferring the viral load that can be collected when the test substrate device is in use. The expected device sensitivity can be determined based on the range of pool size and density.

[0111] The viral load and the number of layers in the test substrate can be determined using the following equation: f(Ln) = VD of a single layer × Ln. Not bound by theory, diminishing returns may exist as more layers need to be built into the function, since exhausted air may not reach the inner layers efficiently. Some devices may only require 1-5 layers to screen smaller pools, while others with more layers may be useful for screening larger pools.

[0112] The range of viral load collection capabilities of the test substrate can also be determined as a function of wear time. Embodiments of the device can be tested at two or more time points. Viral load can be quantified based on the amount of time spent wearing each embodiment.

[0113] Example

[0114] In this example, the use of swabs to collect samples for COVID-19 testing is compared to testing using a face shield insert. Patients presumed to be COVID-19 positive wore a face shield containing the insert for 30 to 60 minutes. After removing the device from the face shield, the test substrate was removed and analyzed by RT-PCR. Simultaneously, nasal swabs were swabbed from the patient and also tested with RT-PCR. The results of the nasal swabs and the face shield insert are compared as summarized in Table 1.

[0115] Table 1. Clinical results of RT-PCR test.

[0116]

[0117] PA224132C

[0118]

[0119] The compositions, apparatus, and methods disclosed herein allow for the rapid and large-scale identification of pathogen infection in populations of individuals or on inanimate surfaces that have come into contact with individuals and / or populations of individuals in a single test. If a positive test result is obtained in a pooled sample, individuals or subpopulations of individuals in the pooled sample can be retested individually to identify one or more infected individuals. The method can also identify individuals exposed through contact with clearly identified infected individuals(s). Subsequent steps can then be taken to isolate both the positive infected individual and the person who came into contact with the positive infected individual to prevent the spread of infection as much as possible. The compositions, apparatus, and methods disclosed herein also reduce the costs associated with individual testing of populations because 1) a single test can be performed on a population of individuals, and 2) relatively inexpensive inserts can be used to allow for reusable masks. Performing a single test on individuals, populations of individuals, and / or subpopulations of individuals reduces the time spent testing multiple individuals individually. The methods disclosed herein also reduce the amount of reagents required to perform a single test compared to the amount required to test multiple individuals individually. Furthermore, the method disclosed herein allows for limited-invasive testing to eliminate the discomfort associated with conventional serological or nasal testing methods.

Claims

1. A mask insert comprising a first layer, a test substrate for capturing an analyte in an air sample taken from a subject, and a second layer, wherein, The first layer and the second layer form an outer layer that substantially surrounds the test substrate, wherein the first layer and the second layer form an overlapping area that does not overlap with the test substrate, and wherein the overlapping areas of the first layer and the second layer combine to form a bonding area and reduce airflow through the mask insert in the bonding area and direct airflow toward the test substrate, wherein at least a portion of the test substrate is bonded to at least one of the first layer and the second layer, and wherein the test substrate is configured to be separated from at least a portion of the outer layer, and wherein the mask insert is configured to attach to the surface of the mask.

2. The mask insert of claim 1, further comprising a fastener located on the surface of at least one of the first layer or the second layer and configured to attach the mask insert to the surface of the mask.

3. The mask insert as claimed in claim 1, wherein the mask insert further includes a perforation portion in the outer layer, near one end of the mask insert.

4. The mask insert as claimed in claim 3, wherein the mask insert further includes a notch extending co-linearly with the perforation portion.

5. The mask insert as claimed in claim 1, wherein, The test substrate includes materials selected from the group consisting of synthetic fibers, natural fibers, and combinations thereof.

6. The mask insert as claimed in claim 5, wherein, The test substrate includes materials selected from the group consisting of hydrophobic fibers, hydrophilic fibers, and combinations thereof.

7. The mask insert as claimed in claim 6, wherein, The hydrophobic fiber is selected from the group consisting of: polypropylene, polylactone, poly-L-lactic acid, polyglycolic acid, co-alkyl acrylate, polybutadiene, polyethylene, polystyrene, polyacrylonitrile, polyethylene terephthalate, polysulfone, polycarbonate, polyvinyl chloride, and combinations thereof.

8. The mask insert as claimed in claim 6 or 7, wherein, The hydrophobic fiber is polycaprolactone.

9. The mask insert as claimed in claim 6, wherein, The hydrophilic fiber is selected from the group consisting of: linear polyethyleneimine, cellulose, cellulose acetate, poly(hydroxyethyl methacrylate), polyethylene oxide, polyvinylpyrrolidone, polyacrylic acid, polyethylene glycol, polyvinyl alcohol, polyvinyl acetate, polyacrylamide, protein, polystyrene sulfonate, and combinations thereof.

10. The mask insert as claimed in claim 5, wherein, The test substrate comprises materials selected from the group consisting of: acrylonitrile / butadiene copolymer, cellulose, cellulose acetate, chitosan, nylon, polyacrylic acid, polyvinyl chloride, polydimethylsiloxane, polyetherimide, polyethersulfone, ethyl acrylate, polyethyl vinyl acetate, poly(ethyl-co-vinyl acetate), polyethylene oxide, polyethylene terephthalate, poly(lactic-co-glycolic acid), polymethacrylate, polymethyl methacrylate, polymethyl methacrylate, polymethylstyrene, polystyrene sulfonate, polystyrene sulfonyl fluoride, poly(styrene-co-acrylonitrile), poly(styrene-co-butadiene), poly(styrene-co-divinylbenzene), polyvinyl acetate, and polyethylene. Alcohols, polyvinyl chloride, polyvinylidene fluoride, polyacrylamide, polyacrylonitrile, polyamic acid, polyaniline, polybenzimidazole, polycaprolactone, polycarbonate, polydimethylsiloxane-co-ethylene oxide, polyether ether ketone, polyethylene, polyethyleneimine, polyimide, polyisoprene, polylactide, polypropylene, polystyrene, polysulfone, polyurethane, polyvinylpyrrolidone, protein, SEBS copolymer, styrene / isoprene copolymer, polystyrene-blended-polyvinylmethyl ether, polyhydroxypropyl methacrylate-blended-polyvinylpyrrolidone, polyhydroxybutyrate-blended-ethylene oxide, polyester-blended-polyhydroxyethyl methacrylate, polyhydroxystyrene-blended-ethylene oxide, and combinations thereof.

11. The mask insert as claimed in claim 5 or 10, wherein, The test substrate comprises materials selected from the group consisting of collagen, fibroinogen, fibronectin, and combinations thereof.

12. The mask insert as claimed in claim 5 or 10, wherein, The test substrate comprises materials selected from the group consisting of: polyvinylidene fluoride-blended-polymethyl methacrylate, polymethyl methacrylate-blended-polyethylene oxide, protein-blended-polyethylene oxide, polylactide-blended-polyvinylpyrrolidone, and polyethylene oxide-blended-polymethyl methacrylate.

13. The mask insert as claimed in claim 5, wherein, The test substrate includes polystyrene-blended polyester.

14. The mask insert as claimed in claim 1, wherein, The test substrate includes electrets.

15. The mask insert as claimed in claim 1, wherein, The first layer and the second layer comprise synthetic fibers, natural fibers, and combinations thereof.

16. The mask insert as claimed in claim 1, wherein the mask insert further comprises a measuring unit selected from the group consisting of a vertical flow measuring unit and a side flow measuring unit.

17. The mask insert of claim 1, wherein the mask insert further comprises a buffer dispensing device.

18. A system for detecting analytes in air samples obtained from a subject, the system comprising: A mask insert comprising a first layer, a test substrate, and a second layer, the test substrate being used to capture analytes from an air sample obtained from a subject, wherein the first layer and the second layer form an outer layer substantially surrounding the test substrate, wherein the first layer and the second layer form overlapping regions that do not overlap with the test substrate, and wherein the overlapping regions of the first layer and the second layer combine to form a binding region and reduce airflow through the mask insert in the binding region and direct airflow toward the test substrate, wherein at least a portion of the test substrate is bound to at least one of the first layer and the second layer, and wherein the test substrate is configured to be separated from at least a portion of the outer layer; and A face mask, which is worn by the subject. The mask insert is attached to the surface of the mask.

19. The system of claim 18, wherein, The mask insert is connected to one of the vertical flow measuring unit and the lateral flow measuring unit.

20. The system of claim 18, wherein, The mask insert also includes a buffer dispensing device.

21. The system of claim 18, wherein, The test substrates include synthetic fibers, natural fibers, and combinations thereof.

22. The system of claim 19, wherein, The test substrate includes electrets.

23. The system of claim 18, wherein, The first layer and the second layer comprise synthetic fibers, natural fibers, and combinations thereof.

Images