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

Abstract

A novel mask insert and system for detecting an analyte in an air sample obtained from a subject is provided. Disclosed herein are compositions, devices, and methods for detecting an analyte in an air sample obtained from a subject. The compositions can be contained on an inanimate surface, such as a mask to be worn by an individual, having a test substrate for capturing the analyte.

Description

Cross - reference to related applications

[0001]

[0001] This application claims the benefit and priority of U.S. Provisional Patent Application No. 63 / 029,974, filed on May 26, 2020, which is incorporated herein by reference in its entirety.

Background Art

[0002]

[0002] The present disclosure generally relates to medicine, as well as to industry and environmental hygiene. More specifically, the present disclosure is directed to compositions, devices, and methods for analytes in air samples obtained from an individual.

[0003]

[0003] Medical monitoring and diagnosis involve sample collection and analysis. Sample collection can include invasive and non - invasive methods. Invasive sample collection methods can include procedures such as surgery and blood sampling that can cause pain, discomfort, and stress to the patient. A milder form of invasive sample collection method can include swabbing. Devices used in invasive sample collection methods can also be expensive and require aseptic handling to prevent sample contamination. Non - invasive sample collection methods can be advantageous over invasive sample collection methods due to the reduction of pain and discomfort in the individual during sample collection.

[0004]

[0004] The rapid detection and identification of pathogens is important for the prevention and monitoring of the presence and spread of infections, for example, the prevention and mitigation of the spread of a pandemic on a global scale. Tests are administered to detect the presence of pathogens on surfaces and in individuals so that appropriate diagnosis, and further testing and / or treatment, can be administered to the individual. It is also possible to administer tests to an individual to detect circulating antibodies against pathogens that indicate that the individual has been previously exposed to the pathogen and has generated an immune response to eradicate the infection.

[0005]

[0005] The detection and identification of analytes, such as biomarkers, can aid in the diagnosis and / or progression of disease, determine the effectiveness and / or response to treatment, and allow for correction and / or adjustment of the dosage of therapeutic agents. The detection of analytes can also be used to detect environmental exposure to potentially harmful chemicals and drug use, such as illegal drug use. The detection of biomarkers can also provide information about the internal physiological state, such as disease, metabolic state, toxins, and the presence of certain chemicals.

[0006]

[0006] Current trials cannot meet the social needs during a pandemic, such as the COVID-19 pandemic. For example, during the COVID-19 pandemic, there was a period when approximately 500,000 trials were conducted per day when more than 20 million trials were requested per day, which demonstrates a trial scale gap. Current trials also suffer from drawbacks and difficulties such as scaling, sample collection and extraction, trial costs, pain associated with sample collection, risk of transmission, trial invalidation (e.g., temperature measurement), and failure to test asymptomatic individuals. Current trials also miss the basic measurement of individuals who are transmitting the disease.

[0007]

[0007] Current tests using nasal swabs for sample collection suffer from limitations in swab supply, sample collection can be painful and uncomfortable, and require collection by skilled healthcare workers. Collection of samples, such as saliva, sputum, drool, lavage, and other oral fluid samples, may require specialized equipment and further processing to separate the analyte from the oral fluid. Regardless of the sample collection method used, the obtained sample may begin to decompose, and sample collection reagents may suffer supply chain delays. The sample then has to be removed or separated from the collection device, which can be inefficient and affect the sensitivity of the test used to analyze the sample. Collection devices for saliva and nasal samples add complexity and are susceptible to contamination of the underlying sample collection matrix; that is, saliva and nasal swabs contain many chemicals that need to be removed before extraction of the target analyte. Proteases and RNases can also decompose the target analyte after extraction. The ratio of the surface area of ​​the recovered material to the required elution and extraction buffers affects the concentration of the target analyte. Higher concentrations of the target analyte result in higher test sensitivity. Identifying the optimal material for forming the factors and performance characteristics provides maximum test sensitivity.

[0008]

[0008] Therefore, there is a need to develop compositions for testing surfaces, groups and individuals of an analyte, devices containing the compositions, and methods for using the compositions. Testing exhaled bioaerosols inexpensively worldwide is a critical and unmet need in the market. [Overview of the project]

[0009]

[0009] In one embodiment, the disclosure relates to a mask insert comprising a first layer, a test substrate for capturing an analyte in an air sample obtained from a subject, and a second layer, wherein the first and second layers form an outer layer substantially surrounding the test substrate, the overlapping regions of the first and second layers are joined, at least a portion of the test substrate is joined to at least one of the first and second layers, and the test substrate is configured to be separated from at least a portion of the outer layer.

[0010]

[0010] In one embodiment, the present disclosure relates to a system comprising a mask insert comprising a first layer, a test substrate for capturing an analyte in an air sample obtained from a subject, and a second layer, wherein the first and second layers form an outer layer substantially surrounding the test substrate, the overlapping regions of the first and second layers are joined, at least a portion of the test substrate is joined to at least one of the first and second layers, and the test substrate is configured to be separated from at least a portion of the outer layer; and a mask worn by a user, wherein the mask insert is connected to the surface of the mask.

[0011]

[0011] In another embodiment, the present disclosure relates to a method for detecting the presence of an analyte in an air sample obtained from a subject, comprising: recovering a test substrate attached by the subject from the solid; and analyzing the test substrate for the analyte.

[0012]

[0012] The present disclosure will be better understood in consideration of the following detailed description, and features, aspects and advantages other than those described above will become apparent. Such a detailed description is referenced to the following drawings. [Brief explanation of the drawing]

[0013] [Figure 1] This is a description showing a front view of one embodiment of a mask insert. [Figure 2]This description shows a front view of one embodiment of a mask insert illustrating the separation of a portion of the outer layer to expose a test substrate. The patient or healthcare provider grasps the outer layer at each end of the mask insert and pulls it outward in the direction of the large arrows (tensile force indicated by the large arrows). The grasping portion of the device may be asymmetrical. The grasping portion on the right side of the description grasps both the outer layer and the inner test substrate. The grasping portion on the left side of the description grasps only the outer layer. By grasping the ends and pulling outward, the perforations allow the left outer protective layer to be removed while the protective layer maintains its hold on the inner test substrate on the right side. After removing a portion of the outer layer, the patient or healthcare provider holds the outer protective layer at the grasping portion on the right side of the description to protect the test substrate from manual contact that may contain RNases, proteases, and other contaminants. The patient or healthcare provider can then place the recovered substrate in a test vial. [Figure 3] This is a description showing the front and side views of the mask insert. The side view shows the stacking and positioning of the micro-hooks for attaching the mask insert to the mask. [Figure 4] This is a description of a mask insert having buffer wells and an integrated vertical flow assay. [Figure 5] This is a description of a mask insert that has a buffer dispensing device and an integrated vertical flow assay. [Figure 6] This description shows a mask insert (with the outer layer removed) with a buffer dispensing device and an integrated vertical flow assay. A multiplex vertical flow assay platform is also shown. [Figure 7] This is a description of a mask insert (with the outer layer removed) that has three buffer dispensing devices and an integrated vertical flow multiplex assay. [Figure 8] This is a description of a mask insert (with the outer layer removed) that has three buffer dispensing devices and an integrated vertical flow multiplex assay. [Figure 9A]This is a description of a mask insert (with the outer layer removed) that has three buffer dispensing plunger devices and an integrated vertical flow multiplex assay. [Figure 9B] This is a description showing a magnified view of three buffer dispensing plunger devices. [Figure 10] This is a description of a mask insert (with an outer layer in a fixed position) that has buffer wells and an integrated lateral flow assay. [Figure 11] This is a description of a mask insert (with a fixed outer layer) that has a buffer dispensing device and an integrated lateral flow assay. [Figure 12] This is a description of a mask insert (with the outer layer removed) that has a buffer dispensing device and an integrated lateral flow assay. [Figure 13A] An exemplary embodiment of a test substrate illustrating the tabs is shown. Figure 13A shows an exemplary embodiment of a test substrate having four tab portions and a label portion. The test substrate may have perforations parallel to the top edge (and below the label portion) and between the individual tabs so that the tabs can be removed independently from the test substrate. [Figure 13B] An exemplary embodiment of a test substrate illustrating the tabs is shown. Figure 13B shows an exemplary embodiment of a test substrate having four tab portions and a label portion. The tab portions are separated by gaps but are attached to the label portion. The test substrate may have perforations parallel to the top edge (and below the label portion) and between the individual tabs so that the tabs can be removed independently from the test substrate. [Figure 13C] An exemplary embodiment of a test substrate illustrating the tabs is shown. Figure 13C shows an exemplary embodiment of a test substrate having four non-connected tab portions. [Figure 14] Figure 13 shows an embodiment of a face mask worn by the user. [Figure 15]An exemplary embodiment of a multilayer test substrate is shown having an inner test substrate made of a material with four tab portions, fully protected and having a high surface area that can be removed from the outer protective layer. [Figure 16] A flow diagram of the extraction and amplification of RNA from the test substrate is shown.

Best Mode for Carrying Out the Invention

[0014]

[0030] The present disclosure generally relates to compositions, devices, and methods for testing inanimate surfaces, groups, and individuals for pathogen infection. In particular, the present disclosure is directed to devices for detecting analytes in air samples exhaled by a subject.

[0015]

[0031] In one aspect, the present disclosure is directed to a mask insert. Referring to FIGS. 1-3, the mask insert includes a first outer layer, a test substrate, and a second outer layer. One of the first layer or the second layer is oriented (proximally) towards the face of the subject, and the other layer is connected (attached) to the inner surface of the mask worn by the subject.

[0016]

[0032] As illustrated in Figures 1-3, the first and second layers are wider and longer than the test substrate (indicated by hash marks, with its outer boundary being a dashed line), and together form the outer layer. The outer ends of the first and second layers that overlap with each other but do not overlap with the test substrate (indicated as diagonal lines " / ") are joined together to form an envelope or sleeve that surrounds the test substrate. The side view shown in Figure 3 illustrates how the inner test substrate is slightly smaller in dimensions than the outer layer. Figure 3 also illustrates the location of fasteners (e.g., micro-hooks in the embodiment shown in Figure 3) on the surface of the outer layer that are away from the face of the subject and function to attach the mask insert to the mask that is more fitted to the subject. The overlapping ends of the first and second layers are also joined together (referred to as "pinching areas" in Figures 1 and 2). The joining of the ends of the first and second layers also creates a barrier that blocks or substantially reduces air from moving through the first and second layers and around the periphery of the test substrate. Therefore, the bonding of the first and second layers directs the airflow towards the test substrate. The outer layer also functions as a protective layer for the test substrate, allowing the mask insert to be handled and manipulated without the test substrate coming into direct contact with it. The outer layer contains material for capturing bioaerosols smaller than 100 microns by providing a bioaerosol bandpass. The outer layer contains an extremely low pressure-drop material that allows bioaerosols to flow easily and reach the test substrate. The outer layer allows bioaerosols to pass through while preventing macrocontaminants that can decompose the analyte, such as RNases and proteases, from coming into contact with the inner test substrate. The mask insert acts like a bandpass filter to allow larger bioaerosols to pass through the outer layer while capturing the analyte with the test substrate.

[0017]

[0033] The ends of the mask insert have micro-hooks or adhesive strips along the ends, which allow the mask insert device to adhere to any face cover material and allow the device to be easily removed without damaging the face cover.

[0018]

[0034] The outer layer has perforations along the lines, so that if the user grasps both ends and pulls the outer layer outwards, a portion of the outer layer can be torn off and discarded. This exposes the inner capturing substrate while the user still holds the outer substrate on one end without contaminating the capturing substrate. Currently, the user can place the inner substrate in a test vial.

[0019]

[0035] The first and second layers may be made from the same or different materials. Suitable materials for making the first and second layers (collectively the outer layer) include, for example, spunbond polypropylene, spunbond polyester, meltblown fibers, and carded nonwoven fibers. Particularly suitable materials for making the first and second layers (collectively the outer layer) include 0.50 oz to 0.75 oz spunbond nonwoven coverweb material (commercially 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 discharged by the subject to pass through the first layer, the test substrate, and the second layer. The material of the first layer, which is close to the mouth and nose of the subject, 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 over it (above) but also to capture analytes contained in the air ejected by the subject. A second layer, located away from the subject's face and close to the mask surface, also allows air to escape from the mask insert device. Suitable fiber sizes for the material forming the outer layer have an average fiber diameter in the range of approximately 5 micrometers to approximately 25 micrometers, for example, in the range of approximately 10 micrometers to approximately 20 micrometers.

[0020]

[0036] As illustrated in Figures 1-3, the mask insert preferably includes a notch in the outer layer adjacent to one end of the mask insert, which allows for easier separation of a portion of the outer layer. As similarly illustrated in Figure 1, the first and second layers include perforations that allow for easier separation of a portion of the outer layer from the test substrate. Preferably, the notch is made in the outer substrate, and the portion ends at the start of the perforation. As further illustrated in Figure 2, the separation of a portion of the outer layer exposes the test substrate. When both ends of the mask insert are held and pulled outward, a portion of the outer layer is pulled away, exposing the test substrate contained within the outer layer, while allowing a person to still grasp the end of the mask insert with the remaining portion without contaminating the test substrate. The test substrate can then be processed to detect the analyte captured on the test substrate. The portion of the mask insert that provided the sleeve containing the test substrate can be discarded after the test substrate is removed. This allows for easy cutting of the outer substrate layer from the inner test substrate while the user still holds one end.

[0021]

[0037] In another embodiment, the disclosure relates to a mask insert incorporating an integrated vertical flow assay (VFA). Figure 4 shows one embodiment of a mask insert having an integrated vertical flow assay. As described in Figure 4, the buffer well is located adjacent to the notches and perforations for separation of a portion of the outer layer as described herein. The well allows the buffer added to the well to come into contact with a portion of the inner test substrate. By capillary action, the buffer flows toward the vertical flow assay located away from the buffer well. If a charged material, such as an electret, is used as the test substrate, the buffer depletes 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 so that the flow is toward the VFA. The test substrate is in contact with the VFA, allowing the buffer containing the analyte to be transported toward the VFA. The buffer may also include other biomarkers that can function as a positive control and / or negative control and can also be detected by the VFA. The VFA portion of the mask insert may include a display "window" to allow observation of the VFA results.

[0022]

[0038] Figure 5 shows one embodiment of a mask insert having an integrated vertical flow assay with a buffer dispensing device located adjacent to the notches and perforations shown in Figure 5. The buffer dispensing device provides a replacement for or addition to the buffer well (shown in Figure 4). The buffer dispensing device may be an ampoule embedded inside the mask insert. When pressure is applied by a person in the mask insert where the buffer dispensing device is located, all or part of the buffer dispensing device ruptures, releasing the buffer contained within it. Through capillary action, the buffer flows toward the vertical flow assay located away from the buffer dispensing device. If a charged material, such as an electret, is used as the test substrate, the buffer depletes 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 so that the flow is toward the VFA. The test substrate is in contact with the VFA, allowing the buffer containing the analyte to be transported toward the VFA. The buffer solution may also contain other biomarkers that function as positive and / or negative controls and can be detected by the VFA. The VFA portion of the mask insert may include a display "window" to allow observation of the VFA results.

[0023]

[0039] Figure 6 shows another embodiment of the mask insert having a removable outer layer, as described in Figure 5. Figure 6 shows a VFA for single analyte detection and a VFA for performing multiplex testing. Removal of 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 further flowing into the material forming the outer layer.

[0024]

[0040] In another embodiment, the mask insert may include two or more buffer dispensing devices. Figure 7 shows an exemplary embodiment of a mask insert including three buffer dispensing devices. The embodiment described in Figure 7 also includes a removable outer layer. The buffer from the buffer dispensing devices flows toward the VFA, but barriers and / or occluding non-porous spacing (or separations) in the test substrate allow for independent elution and extraction of analytes from the test substrate "lanes" and prevent cross-flow of the buffer into other "lanes" of the test substrate intended to detect different analytes. Via capillary action, the buffer flows toward the vertical flow assay located away from the buffer dispensing devices. If a charged material, such as an electret, is used as the test substrate, the buffer depletes 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 so that the flow is toward the VFA. The removal of 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 further flowing into the material forming the outer layer. The test substrate is in contact with the VFA, allowing the buffer containing the analyte to be transported to the VFA. The buffer may also contain other biomarkers that can function as positive and / or negative controls and can be detected by the VFA. The VFA portion of the mask insert may include a display "window" to allow observation of the VFA results. Figure 7 also shows an exemplary embodiment for a multiplex test.

[0025]

[0041] Figure 8 shows another exemplary embodiment of a mask insert including three buffer dispensing devices. The embodiment shown in Figure 8 also includes perforations adjacent to the buffer dispensing devices and adjacent to the VFA to allow removal of only the outer layer surrounding the central portion of the test substrate. The buffer from the buffer dispensing devices flows toward the VFA, but barriers and / or spacing (or separations) in the test substrate prevent each of the buffers from cross-flowing into the test substrate. Through capillary action, the buffer flows toward the vertical flow assay located away from the buffer dispensing devices. If a charged material, such as an electret, is used as the test substrate, the buffer depletes the charge of the electret material as it flows toward the VFA, thereby transporting the analyte to the VFA. Removal of the outer layer of the mask insert between the buffer dispensing devices and the VFA allows the buffer to flow toward the test substrate without further 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 so that the flow is toward the VFA. The test substrate is in contact with the VFA, allowing the buffer containing the analyte to be transported to the VFA. The buffer may also contain other biomarkers that can function as positive and / or negative controls and can be detected by the VFA. The VFA portion of the mask insert may include a display "window" to allow observation of the VFA results. Figure 8 also shows an exemplary embodiment of a multiplex test.

[0026]

[0042] In another embodiment, the mask insert includes an integrated vertical flow assay and at least one integrated delivery plunger for introducing the buffer into the test substrate. Figure 9A shows an exemplary embodiment having three integrated delivery plungers. Figure 9B shows the delivery plungers. The vertical bar on the right is a locking mechanism that can be flipped up to allow the plunger on the left to press, thereby discharging the buffer from the plunger cavity onto the test substrate.

[0027]

[0043] In another embodiment, the mask insert includes an integrated lateral flow assay. Figure 10 shows an exemplary embodiment of a mask insert having an integrated lateral flow assay (LFA). The embodiment in Figure 10 includes a well for introducing a buffer to extract and / or elute the analyte and transport the analyte from the test substrate toward the lateral flow assay sample pad. As shown in Figure 10, the test substrate is in contact with the sample pad of the LFA. The LFA may be integrated with the mask insert in the form of a standard LFA cassette or as a laminated LFA. The embodiment shown in Figure 10 may also utilize multiplex testing.

[0028]

[0044] In another embodiment shown in Figure 11, the mask insert comprises an integrated lateral flow assay further comprising a buffer dispensing device (ampoule) for introducing buffer to extract and / or elute the analyte and transporting the analyte from the test substrate toward the lateral flow assay sample pad. As discussed herein, pressure applied to the buffer dispensing device releases the buffer, which flows through the test substrate via capillary action toward the LFA. As shown in Figure 11, the test substrate is in contact with the sample pad of the LFA. The LFA may be integrated with the mask insert in the form of a standard LFA cassette or as a laminated LFA. The embodiment shown in Figure 11 may also utilize a multiplex assay.

[0029]

[0045] In another exemplary embodiment shown in Figure 12, the mask insert comprises an integrated lateral flow assay further comprising a buffer dispensing device (ampoule) for introducing a buffer to extract and / or elute the analyte and transport the analyte from the test substrate toward a lateral flow assay sample pad. As discussed herein, pressure applied to the buffer dispensing device releases the buffer, which flows through the test substrate via capillary action toward the LFA. Removal of the outer layer of the mask insert between the buffer dispensing device and the LFA allows the buffer to flow through the test substrate without further flowing into the material forming the outer layer. As shown in Figure 12, the test substrate is in contact with the sample pad of the LFA. The LFA may be integrated with the mask insert in the form of a standard LFA cassette or as a laminated LFA. The embodiment shown in Figure 12 may also utilize a multiplex assay.

[0030]

[0046] The test substrate is designed to capture analytes in an air sample from the target. In some embodiments, the test substrate has a pore size smaller than the size of the analyte. In other embodiments, the test substrate is made from a material that attracts the analyte. In some embodiments, the test substrate has a pore size smaller than the size of the analyte and is made from a material that attracts the analyte. Pore sizes for capturing analytes are less than 100 microns. Particularly suitable pore sizes for capturing analytes, such as pathogens, are less than 75 microns, less than 50 microns, and less than 25 microns. The material can be modified to target specific analytes while minimizing pressure drop. For example, capturing COVID may require a higher pressure drop because COVID is smaller and therefore requires a higher density material. TB can be captured with similar efficiency using a lower density material, and therefore a material with lower pressure drop and higher throughput. Exemplary analytes intended to be captured by the test substrate of the mask insert include COVID-19 (approximately 100 nm), influenza (approximately 80–120 nm); Mycobacterium (approximately 7 microns); and other bioaerosols containing respiratory pathogens (less than 10 microns). As described herein, the test substrate material can be treated to "attract" the analyte to the test substrate (e.g., by imparting an electric charge to the material). Thus, for example, a lower-density material can be treated to attract COVID to the test substrate compared to an untreated material.

[0031]

[0047] The mask insert is connected (attached) to the inner surface of the mask. The mask insert is connected to the mask surface by adhesive and other fasteners, such as hook-and-loop fasteners. Preferably, the mask insert includes micro-hooks that allow the mask insert to adhere to the inner surface of the mask to which it is attached. Other connection mechanisms are also acceptable. For example, the mask insert can be attached to the mask surface using adhesive. Generally, a non-permanent adhesive is preferred so that the mask insert can be removed from the mask surface.

[0032]

[0048] Air is expelled through breathing, coughing, sneezing, speaking, and combinations thereof, depending on the subject.

[0033]

[0049] In one embodiment, the mask insert is reversibly attached to the mask worn by the 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.

[0034]

[0050] In another embodiment, the mask insert is semi-permanently attached to the mask worn by the user. In this embodiment, the mask insert can be separated from the mask after it has been attached to the mask surface, but it is not intended to be reattached to the mask insert after it has been removed from the mask surface.

[0035]

[0051] In another embodiment, the mask insert is permanently attached to the mask worn by the user. In this embodiment, the mask insert is not intended to be separated from the mask once it has adhered to the mask surface.

[0036]

[0052] Generally, a mask insert (whether reversibly or permanently connected) is positioned on the inner surface of the mask where air exhaled by the user through normal breathing, labored breathing, coughing, sneezing, and any combination thereof causes airflow to the mask insert.

[0037]

[0053] As used herein, the substrate to be tested (also interchangeably referred to herein as “test substrate” and “recovered substrate”) may include any inanimate surface that may come into contact with an individual or population that may contain pathogens on its external surface. For example, in one embodiment, the substrate to be tested may be an entire mask or a mask from which a portion of the mask (i.e., the test substrate) has been removed. The substrate may also include a removable mask insert. In some embodiments, the mask insert can be reused by cleaning it and returning it to the mask.

[0038]

[0054] In some embodiments, the test substrate is processed 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 produces 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) in which the test substrate dissolves. In another embodiment, a reagent is applied to the test substrate. In another embodiment, the reagent is brought into contact with the test substrate, which is drawn into the test substrate by capillary action.

[0039]

[0055] The test substrate may preferably be made of synthetic fibers, natural fibers, or combinations thereof. The fibers used to form the layer may be hydrophobic fibers, hydrophilic fibers, or 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), proteins, 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-acrylonitrile), Examples include poly(styrene-co-butadiene), poly(styrene-co-divinylbenzene), poly(vinyl acetate), poly(vinyl alcohol), poly(vinyl chloride), poly(vinylidene fluoride), polyacrylamide, polyacrylonitrile, polyamine 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, raw silk, styrene / isoprene copolymer, and combinations thereof.Polymer blends, such as 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.

[0040]

[0056] Another suitable layer of the test substrate may be an electret (e.g., thermal electrets and fibrillated electret films). An electret is a dielectric material having a pseudo-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 dipoles in the material in a favorable direction. After heating, the material can be "frozen" to retain the position of its electric dipoles for a long period of time. Suitable materials for preparing electrets include, for example, materials that can currently be used to produce thermal electrets, such as organic materials, e.g., ebonite, naphthalene, polymethyl methacrylate, and many polymers, as well as inorganic materials, e.g., sulfur, quartz, glass, steatite, and some ceramics. Electret fiber films are particularly preferred. Polyvinylidene fluoride (PVDF) / polytetrafluoroethylene (PTFE) NP electret nanofiber films can be formed by electrospinning. The fibrillated electret film described in Van Turnhout (U.S. Patent No. 3,998,916) is also suitable.

[0041]

[0057] Particularly suitable test substrates are charged materials that attract analytes contained in the air sample (e.g., polypropylene, polylactic acid, and electrets). Therefore, although the analytes contained in the air are not necessarily captured by the test substrate due to the size difference between the test substrate and the analytes, they are attracted to the test substrate.

[0042]

[0058] Embodiments using polypropylene as the test substrate advantageously provide an inert substrate that allows biological materials, such as pathogens, to remain stable for extended periods, for example, when the mask insert and / or test substrate require transport for testing. The test substrate may also include coatings and chemical treatments that allow for easier elution of the analyte. The wetting ability of the test substrate material can be increased by using a good low surface energy material, thus impregnating the material and thus requiring less buffer to concentrate the analyte in the elution buffer. Hydrophobic polypropylene has the advantage of readily discharging and allowing elution of virions from the nonwoven fabric with a simple application of a buffer, for example, 0.1% Triton X-100 to perform crude extraction for a certain pathogen, e.g., SARS-CoV-2.

[0043]

[0059] The test substrate material can also be treated to impart an electric charge to increase or decrease the hydrophilicity of the material, increase or decrease the hydrophobicity of the material, or a combination thereof.

[0044]

[0060] Suitable test substrate materials include materials that are soluble and / or soluble in liquids. For example, cellulose acetate nanofibers that dissolve upon contact with a liquid. It should be understood that the entire test substrate and / or a portion thereof (e.g., the recovered substrate) may be soluble or soluble. Advantageously, the substrate itself may dissolve completely, releasing all analyte material into the eluent without requiring an extraction process. Preferably, the substrate may be prepared such that it dissolves, reacts, stabilizes RNA, or performs some other service detection. Preferably, the substrate may be inert before it is dissolved. Furthermore, or / or, a protective inert layer of the dissolving inner substrate exposes a low-inert, active inner substrate with activity diagnostic properties. For example, the test substrate (e.g., a rod) is placed in an eluent that dissolves the outer coating, releasing the entire amount of viral material. The test reagent then dissolves and interacts with the analyte material.

[0045]

[0061] Fibers may be subnanofibers, nanofibers, microfibers, or combinations thereof, having diameters in the subnanometer to micrometer range. Subnanofibers can be prepared using template-assisted growth, orientation-assisted growth, ligand-controlled growth, and catalyst-guided growth. Subnanometer fibers (also referred to herein as “subnanofibers”) refer to a single dimensional structure with an extremely high aspect ratio having a diameter in the range of several angstroms to tens of angstroms. Ultrathin nanofibers with a diameter of less than 100 nm can be produced by phase separation, self-assembly, the Sea Island method, template synthesis, electrospinning, and bubble-electrospinning. Examples of nanofiber diameter ranges include about 3 nm to less than 1 μm. Examples of microfiber diameter ranges include 1 μm to about 10 μm. Nanofibers and microfibers can be produced using phase separation, self-assembly, the Sea Island method, template synthesis, electrospinning, and bubble-electrospinning, melt-blown, spin-blown, and other methods. The fiber diameter and morphology can be determined using scanning electron microscopy (SEM), transmission electron microscopy (TEM), and atomic force microscopy (AFM). Test substrate fibers may be woven, knitted, crocheted, knotted, co-pressed, interlaced, bonded, co-stapled in the form of sheet webs, or combinations thereof. Layers can be formed using spunbond materials prepared with continuous fibers formed by continuously extruding a polymer through a spinneret to form distinct filaments. The filaments can then be mechanically or pneumatically stretched without interrupting the orientation of the polymer filaments. The continuous filaments can be deposited substantially randomly on a carrier belt to form a web. Layers can also be formed using meltblown fibers. Layers can also be formed using electrospun fibers. As is known to those skilled in the art, electrospinning involves stretching individual polymer chains in a polymer solution through a nozzle / orifice, typically having a very small diameter, under high voltage to form nano- or sub-microscale structures in the form of fibers.

[0046]

[0062] Additives can be included in the test substrate layer. Suitable additives include, for example, antimicrobial additives such as silver-containing antimicrobial agents and antimicrobial polypeptides, analgesic compounds 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, other pharmaceutical compounds, adhesives, fragrances, deodorizing compounds, preservatives, RNase inhibitors, protease inhibitors, and nucleic acids, such as deoxyribonucleic acid, ribonucleic acid, and nucleotide analogs.

[0047]

[0063] Suitable analytes are contained in gases and aerosol droplets in the air discharged by the user. Suitable analytes include microorganisms, chemical substances, proteins, nucleic acids, and combinations thereof. Suitable microorganisms include bacteria and viruses.

[0048]

[0064] Particularly suitable microorganisms include pathogens. The term "pathogen" is used according to its usual meaning to refer to bacteria, viruses, and other microorganisms that directly or indirectly cause disease. Exemplary pathogens include, for example, the genera Yersinia, Klebsiella, Providencia, Erwinia, Enterobacter, Salmonella, Serratia, Aerobacter, Escherichia, Pseudomonas, and Shigella. Vibrio, Aeromonas, Streptococcus, Staphylococcus, Micrococcus, Moraxella, Bacillus, Clostridium, Corynebacterium, Eberthella, Francisella Haemophilus, Bacteroides, Listeria, Erysipelothrix, Acinetobacter, Brucella, Pasteurella, Flavobacterium, Fusobacterium, Streptobacillus (Obacillus), Calymmatobacterium, Legionella, Treponema, Borrelia, Leptospira, Actinomyces, Nocardia, Rickettsia, Micrococcus, Mycobacterium,Genera Neisseria, Campylobacter, pathogenic viruses such as papillomavirus, parvovirus, adenovirus, herpesvirus, vaccine virus, arenavirus, coronavirus, rhinovirus, respiratory syncytial virus, influenza virus, picornavirus, paramyxovirus, reovirus, retrovirus, rhabdovirus, human immunodeficiency virus (HIV), etc., genera Taenia, Hymenolepsis, Diphyllobothrium lobothrium), Echinococcus, Fasciolopsis, Heterophyes, Metagonimus, Clonorchis, Fascio la), Paragonimus, Schistosoma, Enterobius, Trichuris, Ascaris, Ancylostoma, Necator, Wuc hereria, Brugi, Loa, Onchocerca, Dracunculus, Naegleria, Acanthamoeba, Plasmodium, Trypanosoma, Leishmania, Toxoplasma, Entamoeba, Giardia, Isospora pora), Cryptosporidium, Enterocytozoa, Strongyloides, Trichinella, for example, tinea, histoplasmosis, blastomycosis, aspergillosis, cryptococcosis, sporotrichumosis, coccidiodomycosis, paracoccidioidomycosis, mucomycosis, candidiasis, dermatophytosis, protothecasis, pityriasis, mycomas,Examples include fungi of the genus Pneumocystis and combinations thereof that cause paracoccidiodomycosis, pheophomycosis, pseudoalescheririosis, and trichosporonosis.

[0049]

[0065] Particularly suitable chemical substances include ketones, nicotine, cocaine, opioids, marijuana, benzodiazepines, amphetamines, barbiturates, and combinations thereof.

[0050]

[0066] It can also detect proteins, DNA, and RNA.

[0051]

[0067] Suitable masks include any face covering that is worn over or in front of the user's mouth and nasal cavity, from which air is expelled from the user, for example, during exhalation during speaking, coughing, and sneezing. Masks can be secured to the user's face using ties, straps, bands, and combinations thereof. Masks also include face shields. Masks also include bandanas, neck gaiters, scarves, towels, and cloth covers that are positioned over the user's nose and mouth. Thus, the mask inserts of this disclosure can be coupled with any type of face covering so that at least a portion of the air expelled (by breathing, coughing, sneezing, or speaking) passes through the device, and the analytes contained in the air sample can be recovered by the test substrate using the device.

[0052]

[0068] When the analyte is a pathogen, the amount of pathogen particles captured can be expressed as filtration efficiency, minimum efficiency report value evaluation, and particulate performance evaluation. Preferably, the filtration efficiency of the test substrate is in the range of about 60% to about 95%. Preferably, the minimum efficiency report value evaluation is at least 12. Preferably, the particulate performance evaluation is at least 1900.

[0053]

[0069] Any suitable method, such as heat treatment, ultraviolet light irradiation, and cleaning and / or sterilization by other sterilization methods, can be used to clean the mask and / or mask insert. In other embodiments, the mask and / or mask insert is a disposable mask or mask insert. In these embodiments, the mask and / or mask insert is destroyed after removal.

[0054]

[0070] In one embodiment, the disclosure relates to a mask for testing for pathogen infection. The mask comprises a test substrate containing a capture agent for capturing pathogens.

[0055]

[0071] Suitable capture agents include, for example, antibodies that specifically bind to pathogens, ligands that specifically bind to pathogens, and surface molecules to which pathogens are expected to bind in order to infect their host, such as sugars and glycoproteins. The capture reagent may be covalently or noncovalently linked to the test substrate by a linker. Any suitable linker can be used, such as organic molecules, such as polymers or copolymers (e.g., substituted or unsubstituted polyalkylene glycols, e.g., polyethylene glycol), and / or biomolecules, such as bovine serum albumin.

[0056]

[0072] In one embodiment, a mask worn by an individual covers the mouth and nose of the individual, and as a result, pathogens are captured and / or adsorbed by the mask material when the individual breathes, coughs, and / or sneezes while inside the mask. The mask may include ear loops and / or ties for securing the mask to the face of the individual wearer. The mask may include strips of material attached to the mask for use in attaching the mask to the face of the wearer, extending along each of its sides, for providing an improved fluid seal between the periphery of the mask and the wearer's face.

[0057]

[0073] The test substrate may be coated and / or treated to absorb and / or capture pathogens.

[0058]

[0074] The mask and / or the mask insert portion of the mask can be constructed to maximize the surface area of ​​the test substrate. For example, the test substrate may contain particles, such as microparticles, nanoparticles, and beads. The particles can be coated with a scavenging agent.

[0059]

[0075] Preferably, 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. Examples of substrates include 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-resolved chromatography test substrates, chemiluminescence test substrates, and other test substrates.

[0060]

[0076] In embodiments of a mask intended to contain a test substrate, the mask is configured to receive the test substrate. The mask may have perforations, for example, allowing the test substrate to be inserted into the mask at the perforations. In other embodiments, the test substrate may be attached to the mask by means of, for example, hook-and-loop fasteners and adhesives.

[0061]

[0077] The mask insert may also include a tab for labeling the mask insert and for handling the mask without contaminating the test substrate.

[0062]

[0078] As illustrated in Figure 1, perforations in the mask insert, particularly in the test substrate, can be formed to allow the test substrate to be separated into different parts (or tabs). Perforations may be present between each tab. Furthermore, or alternatively, perforations may be parallel to the upper (or lower) end of the test substrate. As shown in Figures 1A and 1B, in addition to perforations between each tab, perforations can be oriented to allow individual tabs to be separated from the label portion, thereby allowing individual tabs to be separated from the test substrate independently of the other tabs of the test substrate, leaving the remaining tabs attached. Furthermore, or alternatively, the test substrate may have perforations along the upper (or lower) end of the test substrate, and the space (or gap) may separate each tab so that each tab is independently connected to the upper or lower end of the test substrate, as shown in Figure 1B. In another embodiment illustrated in Figure 1C, the tabs may be "floating" within the test substrate. For example, the test substrate may have four individual tabs that are not connected but contained in the same plane of the test substrate, rather than a single strip with perforations creating four tabs. The exemplary embodiments described in Figures 1A-1C show four tab portions that can be tested individually. For example, tab 1 can be pooled with other tabs obtained from multiple patients. Tab 2 can then be tested individually. Tab 3 can be used as a control. Tab 4 can be stored for future testing. While Figure 1 illustrates an exemplary embodiment having four tabs, it should be understood that the test substrate may have fewer than four tabs and five or more tabs. Figure 2 illustrates a user-mounted mask having a test substrate with four test tabs.

[0063]

[0079] It should be understood that “tab” can refer to the recovered substrate alone or a separable portion of the test substrate. For example, a test substrate tab may include one or more protective layers having the recovered substrate sandwiched between protective layers (and other layers). In this embodiment, the test substrate tab (i.e., a portion of the test substrate) can be separated from the rest of the test substrate, leaving the other tab portion undamaged. In another embodiment, the recovered substrate tab can be sandwiched between protective layers. In this embodiment, the recovered substrate tab can be removed independently from between the protective layers without (one or more) protective layers and / or without contact with or damage to the rest of the recovered substrate tab. The advantage of the “tooth-shaped” embodiment is that a physician can easily hold the tab and tear along the perforations along the top, which removes both the other and inner layers from a common back, and the inner layer can be easily placed in a container for further testing without contamination, and the outer layer is discarded, etc. A tab may also include multiple layers as described herein with respect to multilayer test substrates. (One or more) tabs may share a common back but be separable (as shown, for example, in Figure 1C). The tabs may have perforations for easy removal and to prevent contamination.

[0064]

[0080] The test substrate may be a multilayer test substrate. A multilayer test substrate has an inner recovery substrate that is entirely protected by an additional layer. The inner recovery layer is made from a material with a high surface area for the recovery of pathogens to be detected. The inner recovery substrate can also be separated from the protective layer. The protective layer and the inner recovery substrate can be made from different materials designed for specific purposes of the layers (e.g., protection and sample recovery). For example, the inner recovery substrate may be made from corrugated double-sided polyester swab material. The inner recovery substrate can be coated with a reagent for holding the recovered pathogen load. The inner recovery substrate can also be coated with a reagent for stabilizing the pathogen material. Figure 3 shows a multilayer test substrate. The protective layer may also function as a porous, flexible transport layer capable of receiving and temporarily storing exhaled droplets before they are transferred and absorbed by one or more lower recovery layers. The protective layer may be an air-permeable contact protective coating. The protective layer located furthest from the user's face may block all airflow, while the protective layer located closest to the user's face may allow airflow.

[0065]

[0081] In some embodiments, (one or more) inner recovery substrates may be separated from the protective layer. In some embodiments, the protective layer may be open to allow removal or exposure of the inner recovery substrate. For example, in an exemplary embodiment having an adjacent protective layer and a separation protective layer, with the inner recovery substrate positioned between the protective layers, the protective layer may be larger in size than the inner recovery substrate. When sandwiched together, the surfaces of the protective layers may be in contact with each other and may be held together using adhesive, press-fit locking zippers, tape, or hinges. Separation of the protective layer allows access to the inner recovery substrate which can be removed for testing. Separation of the protective layer also allows insertion or replacement of unused inner recovery substrate.

[0066]

[0082] Features of multilayer test substrates and multilayer tabs include, for example, that (one or more) outer layers protect inner layers from contamination, (one or more) inner layers may be bifacial, (one or more) inner layers may contain adhesives, (one or more) inner layers may contain stabilizers, (one or more) inner layers may contain test layers, and (one or more) inner layers may be multilayered.

[0067]

[0083] In one embodiment, a multilayer test substrate may comprise one or more reagent layers (also referred to herein as “reagent material” containing analytical reagents). As used herein, “analytical reagent” refers to components used for the detection of pathogens. For example, analytical reagents may include buffer components, salts, dNTPs, oligonucleotide primers, polymerases, reverse transcriptases, and combinations thereof. Analytical reagents may preferably be lyophilized, in liquid form, gel, and combinations thereof. The reagent layer containing the analytical reagent can be separated from other layers of the test substrate by a coating to prevent the reagent from contacting and / or activating the recovery layer until the test substrate is to be processed for analysis. The analytical reagent can be activated, for example, by placing the test substrate (and / or the recovery layer having a soluble layer containing the reagent) in a liquid medium, such as a buffer, e.g., water, so that the coating dissolves and releases the analytical reagent, which can then also dissolve in the buffer. The coating may be meltable, and the temperature can be adjusted so that the coating melts and releases the analytical reagent, forming a mixture in which pathogens can be detected. The reagent layer may contain microparticles and / or beads containing analytical reagents.

[0068]

[0084] In some embodiments, the test substrate and (one or more) inner recovery substrates can be packaged in sterile packaging material to prevent contamination during storage and handling. The packaging material may then be open to allow the test substrate or inner recovery substrate to be placed in the mask and / or with the protective layer.

[0069]

[0085] The multilayer test substrate may include spacers between the individual layers. The spacers may be positioned at the edges of the recovery area to prevent air from flowing through the spacer material. Furthermore, or alternatively, the spacers may be made of a material through which air and pathogen particles can pass.

[0070]

[0086] Multilayer test substrates may include labels (see, for example, Figure 1). The labels may be blank areas on which information can be printed. The labels may be pre-printed and may have machine-readable barcodes. The information contained on the labels can be used to identify test substrates (including individual test substrate tabs), track recovered test substrates, and associate them with users.

[0071]

[0087] In a multilayer test substrate, individual layers may have different pore sizes than other layers. In one exemplary embodiment, the first protective layer may have a pore size that has low filtration efficiency, allowing pathogen-sized particles to freely pass through the protective layer and reach the recovery layer.

[0072]

[0088] In an exemplary embodiment, the test substrate may have a first protective layer made of a single fibrous material having a pore size range that allows pathogen-sized particles to pass freely through the first protective layer, and at least one recovery layer made of a different fibrous material having a pore size smaller than that of the pathogen-sized particles so that the pathogen-sized particles are captured by the recovery layer. Preferably, the recovery layer may be a multilayer laminate itself to increase the virus capture load of each test. If the inner substrate tab can capture the virus density of X, there should be a function of f(Ln) = Ln × virus load of a single layer (where Ln is the number of layers). The reduction in efficiency due to the addition of layers must be understood.

[0073]

[0089] One or more recovery layers can be treated to improve the capture of pathogen particles. In one embodiment, the recovery layer is treated with an adhesive to which pathogen particles adhere. In another embodiment, the recovery material is treated with a capture ligand. In another exemplary embodiment, the recovery material is treated with a combination of adhesive and capture ligand. The capture ligand may be an antibody containing an antibody fragment, an aptamer, a magnetic particle, or other ligand type that can bind to the pathogen of interest. The capture ligand is intended to attract and retain the pathogen so that subsequent analysis can be performed to detect and / or identify the pathogen.

[0074]

[0090] An exemplary multilayer test substrate includes, for example, only a recovery layer without a scavenger and without a protective layer. Another exemplary multilayer test substrate includes a protective layer and a recovery layer. Another exemplary multilayer test substrate includes a protective layer and a recovery layer, and the recovery substrate includes a scavenger (e.g., adhesives, scavenging ligands, and combinations thereof). Another exemplary multilayer test substrate includes a protective layer and multiple recovery layers.

[0075]

[0091] The protective layer may be made of a material that allows for protection of the airflow and the inner recovery layer. The protective layer can be treated or coated with reagents, such as antimicrobial agents, RNase inhibitors, protease inhibitors, preservatives, and combinations thereof. The protective layer also provides, for example, physical protection of the inner recovery substrate from contact. Each layer may have perforations to allow for separation of the tabs.

[0076]

[0092] The orientation of the test substrate can be planar (similar to a paper sheet), coiled, rod-shaped, hollow cylindrical, honeycomb, or a combination thereof. Coiled, rod-shaped, and hollow cylindrical substrates may be available axially or perpendicularly to the breathing direction. Coiled, rod-shaped, and hollow cylindrical substrates can also be easily housed in tubes for storage and / or processing. It should be understood that each of these embodiments provides a three-dimensional structure to improve recovery and allow airflow.

[0077]

[0093] In one embodiment, the disclosure relates to a method for detecting the presence of a pathogen. In one embodiment, the method detects the presence of a pathogen on an inanimate surface that may come into contact with an individual or population. In another embodiment, the method detects the presence of a pathogen infection in a population in which at least one individual has or is suspected to have a pathogen infection. The method comprises: recovering a test substrate attached by each individual in a population from a population; combining the test substrates recovered from each individual to form a pooled sample of test substrates; and analyzing the pooled sample of test substrates for a pathogen, wherein the detection of a pathogen in the pooled sample of test substrates indicates that at least one individual in the population has had close exposure to the pathogen, is infected with the pathogen, or may require further diagnostic testing.

[0078]

[0094] The method may further include independently administering a second test to each individual in the group to identify individuals infected with the pathogen.

[0079]

[0095] In some embodiments, the method is repeated for subgroups or subpools of the first population. For example, the method in this embodiment involves: recovering test substrates attached by each individual in the first population; combining the test substrates recovered from each individual to form a pooled sample of the first test substrates; and analyzing the pooled sample of the first test substrates for a pathogen, wherein the detection of the presence of a pathogen in the pooled sample of the test substrates indicates that at least one individual in the population has had close exposure to the pathogen, is infected with the pathogen, or may require further diagnostic testing; and then the second population The method includes: recovering test substrates attached by each individual in the population, wherein the second population is a subgroup of the first population; combining the test substrates recovered from each individual to form a pooled sample of the second test substrate; and analyzing the pooled sample of the second test substrate for pathogens, wherein the detection of the presence of pathogens in the pooled sample of the test substrate indicates that at least one individual in the population has had close exposure to the pathogen, is infected with the pathogen, or may require further diagnostic testing. The method can be repeated any number of times to identify individuals or subgroups of individuals infected with the pathogen, or material substrates that come into contact with such individuals or subgroups of individuals.

[0080]

[0096] Suitable test substrates are described herein. Preferably, the test substrate is a mask or a part of a mask (e.g., a mask insert) as described herein.

[0081]

[0097] Suitable pathogens to be tested are described herein.

[0082]

[0098] In one embodiment, the Disclosure relates to a method for detecting the presence of a pathogen in an individual having or suspected to have a pathogen infection, the method comprising: recovering a test substrate attached by the individual from the individual; and analyzing the test substrate for a pathogen, wherein the detection of the presence of a pathogen indicates that the individual has had close exposure to the pathogen, is infected with the pathogen, or may require further diagnostic testing.

[0083]

[0099] Suitable test substrates are described herein. Preferably, the test substrate is a mask or a part of a mask as described herein (e.g., a mask insert, a test substrate, a tab).

[0084]

[0100] In some embodiments, the (one or more) pathogens and / or pathogenic material to be tested are extracted (eluted or removed) from the test substrate. In some embodiments, the test substrate can be analyzed directly, for example, in embodiments where the test substrate used (including tabs, inner test substrate layers, etc.) is a soluble test substrate, an evaporative test substrate.

[0085]

[0101] Suitable pathogens to be tested are described herein.

[0086]

[0102] The test substrates and the test substrates used in the methods of this disclosure can be tested directly, or they can be removed from a mask, for example, by cutting the mask and removing a portion of the mask to be tested, or the test substrates in the form of mask inserts can be configured for use in an analyzer. Any analyzer used to test the substrate for the presence of an analyte (e.g., a pathogen in this disclosure) is suitable for use in this disclosure. In one embodiment, the test substrate can be inserted into an analyzer and the pathogen can be detected by the analyzer. In other embodiments, the analyzer may be a point-of-care analyzer, for example, by directly dropping a solution containing the detection reagent onto the test substrate to bring the detection reagent into direct contact with the test substrate. These point-of-care analyzers may be for personal use at home, use in the workplace, etc. The signal output of the analyzer may provide whether or not a pathogen is present 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. Preferably, the test substrate (including tabs) can be placed in an ion mobility spectrometer and tested for pathogens.

[0087]

[0103] As illustrated in Figure 4, the test substrate is adapted to additional sample preparation methods, such as the application of mechanical, chemical, and force (e.g., magnetic force).

[0088]

[0104] Figure 5 shows a flowchart for sample detection using amplification. After recovery, RNA is extracted from the sample and subjected to reverse transcription to create a pool of cDNA. The cDNA is then amplified (e.g., using polymerase chain reaction ("PCR") with primers specific to the pathogen of interest). After amplification, the PCR product can be further analyzed. The analysis may include a simple product / no product result and / or the PCR product can be sequenced. The amount of product can also be used to determine the concentration of the pathogen.

[0089]

[0105] After sample collection via the mask and / or test substrate, the collected sample can be analyzed using commercially available equipment and methods.

[0090]

[0106] The range of virus density and load capacity of the above device embodiment can be determined by utilizing a ratio based on an internally controlled spike test on the mask.

[0091]

[0107] The approximate range of viral load recovered by the embodiment of the test substrate device is assumed to be VD. d1 / VD m1 :VD d2 / VD m2 (In the formula, VD = 1 square millimeter (mm) 2 ) The viral density of the inner substrate normalized based on the viral load per unit; d1 = pathogen spike device in the mask; m1 = pathogen spike mask with the device in the mask; d2 = estimated performance of the device embodiment; m2 = mask recovered from the patient. This can be estimated for infected humans wearing the device under different conditions of use. Once the range of results from the pathogen spike test and recovered masks is determined, the approximate range of recovery potential of the device embodiment can be estimated. In another example, the test substrate device can spike at a range of viral loads and simulate low, average, and high densities to estimate the viral load that can be recovered by the test substrate device in use. Predicted device sensitivity can be determined based on the range of pool size and the range of density.

[0092]

[0108] The virus load and number of layers of the test substrate can be determined using the formula: f(Ln) = VD × Ln of a single layer. Although not theoretically constrained, the return value may decrease if there are many layers that need to be incorporated into the function, as the discharged air may not be able to efficiently reach the inner layers. Some devices may require only 1 to 5 layers to screen smaller pools, while others with more layers are useful for screening larger pools.

[0093]

[0109] The range of viral load recovery capacity of the test substrate can also be determined as a function of the wearing time. The device embodiments can be tested at two or more time points. The viral load can be quantified based on the amount of time each embodiment is worn.

[0094]

[0110] [Examples]

[0111] In this embodiment, the detection of COVID-19 using swabs to collect samples was compared with detection using mask inserts. Patients suspected of being COVID-19 positive wore masks containing mask inserts for 30–60 minutes. After removing the device from the mask, the test substrate was removed and analyzed by RT-PCR. Simultaneously, patients were swabbed with nasal swabs, which were also tested by RT-PCR. The results for nasal swabs and mask inserts were compared as summarized in Table 1.

[0095]

[0112] [Table 1]

[0096]

[0113] The compositions, devices, and methods of this disclosure enable rapid, large-scale identification of pathogen infections in populations or on inanimate surfaces in contact with individuals and / or populations, in a single test. If a positive test result is obtained in a pooled sample, individuals or subgroups of individuals in the pooled sample can be retested separately to identify infected individuals and / or multiple individuals. The methods can also identify individuals exposed through contact with (one or more) infected individuals identified as positive. Subsequent steps can then be taken to isolate both the positive infected individuals and those exposed to them, thereby preventing further spread of infection. The compositions, devices, and methods of this disclosure also reduce the cost associated with individual testing of populations because 1) a single test can be administered to populations, and 2) masks can be reused using relatively inexpensive inserts. Administering a single test to individuals, populations, and / or subgroups of individuals reduces the time required to conduct separate tests on multiple individuals. The methods of this disclosure also reduce the amount of reagents required to conduct a single test compared to the amount of reagents required to test multiple individuals separately. Furthermore, the method of this disclosure enables limited invasive testing and eliminates the discomfort associated with conventional serological or nasal testing methods.

Claims

1. 1. A mask insert comprising: a first layer, a test substrate for capturing an analyte in an air sample obtained from a subject; and a second layer, wherein the first layer and the second layer form an outer layer substantially surrounding the test substrate, an overlapping area of ​​the first layer and the second layer are bonded, at least a portion of the test substrate is bonded to at least one of the first layer and the second layer, the test substrate is configured to be separated from at least a portion of the outer layer, and the mask insert is configured to couple to a surface of a mask.

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

3. 10. The mask insert of claim 1, further comprising perforations in the outer layer proximate an end of the mask insert.

4. A mask insert as described in claim 1, further comprising a notch coextensive with the perforation.

5. 10. The mask insert of claim 1, wherein the test substrate comprises a material selected from the group consisting of synthetic fibers, natural fibers, and combinations thereof.

6. 6. The mask insert of claim 5, wherein the test substrate comprises a material selected from the group consisting of hydrophobic fibers, hydrophilic fibers, and combinations thereof.

7. 7. The mask insert of claim 6, wherein the hydrophobic fibers are selected from the group consisting of polypropylene, polylactone, poly(caprolactone), poly(L-lactic acid), poly(glycolic acid), co-poly(alkyl acrylate), polybutadiene, polyethylene, polystyrene, polyacrylonitrile, polyethylene(terephthalate), polysulfone, polycarbonate, poly(vinyl chloride), and combinations thereof.

8. 7. The mask insert of claim 6, wherein the hydrophilic fibers are selected from the group consisting of 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), proteins, poly(vinylpyrrolidone), poly(styrenesulfonate), and combinations thereof.

9. The test substrate may be any of acrylonitrile / butadiene copolymers, 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-co-glycolic acid), poly(methacrylic acid) salts, poly(methyl methacrylate), ... poly(vinyl methacrylate), poly(methylstyrene), poly(styrenesulfonate), poly(styrenesulfonyl fluoride), poly(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 [0023] The present invention relates to a method for producing a polyester blend comprising the steps of: blending a polyester-co-polyethylene oxide, polyetheretherketone, polyethylene, polyethyleneimine, polyimide, polyisoprene, polylactide, polypropylene, polystyrene, polysulfone, polyurethane, polyvinylpyrrolidone, protein, SEBS copolymer, raw silk, styrene / isoprene copolymer, 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.The mask insert according to claim 5.

10. The mask insert of claim 1 , wherein the test substrate comprises an electret.

11. 10. The mask insert of claim 1, wherein the first layer and the second layer comprise synthetic fibers, natural fibers, and combinations thereof.

12. 10. The mask insert of claim 1, wherein the bond substantially reduces air flow through the bonded area of ​​the mask insert.

13. 10. The mask insert of claim 1, further comprising an assay selected from the group consisting of a vertical flow assay and a lateral flow assay.

14. The mask insert of claim 1, further comprising a buffer dispensing device having an integrated vertical flow assay.

15. 1. A system for detecting an analyte in an air sample obtained from a subject, comprising: a mask insert including a first layer, a test substrate for capturing an analyte in an air sample obtained from a subject, and a second layer, the first layer and the second layer forming an outer layer substantially surrounding the test substrate, an overlapping area of ​​the first layer and the second layer being bonded, at least a portion of the test substrate being bonded to at least one of the first layer and the second layer, and the test substrate being configured to be separated from at least a portion of the outer layer; a mask suitable for being worn by the subject; and Including, The mask insert is coupled to a surface of the mask.

16. The system of claim 15 , wherein the mask insert is coupled to one of a vertical flow assay and a lateral flow assay.

17. The system of claim 15 , wherein the mask insert further comprises a buffer dispensing device with an integrated vertical flow assay.

18. The system of claim 15 , wherein the test substrate comprises synthetic fibers, natural fibers, and combinations thereof.

19. The system of claim 16 , wherein the test substrate comprises an electret.

20. 16. The system of claim 15, wherein the first layer and the second layer comprise synthetic fibers, natural fibers, and combinations thereof.

Images