WO2025240785A1

WO2025240785A1 - Sample collection device and system and a method of collecting and analyzing a sample

Info

Publication number: WO2025240785A1
Application number: PCT/US2025/029631
Authority: WO
Inventors: Michael Wohl; Michael R. Berrigan; Laura R. NERENG; John B. STENDER; Marc K. WAINIONPAA
Original assignee: 3M Innovative Properties Co
Current assignee: 3M Innovative Properties Co
Priority date: 2024-05-16
Filing date: 2025-05-15
Publication date: 2025-11-20
Anticipated expiration: 2026-11-16

Abstract

A sample collection system includes a housing comprising an air inlet constructed to receive an exhalation airflow from a nostril; a porous sample collection media; and an airflow path extending through the porous sample collection media. A sample collection device may include porous sample collection media, a holder, and a sample receiving element. The system may include a liquid reservoir in fluid communication with the porous sample collection media, constructed to direct liquid onto the porous sample collection media. The system may include an assay constructed to receive an eluted sample from the porous sample collection media. A method of collecting and testing a sample includes flowing exhalation air from a nostril through the porous sample collection media to form a captured sample; releasing a metered dose of liquid onto the porous sample collection media to elute the sample onto the assay; and observing a test result on the assay.

Description

SAMPLE COLLECTION DEVICE AND SYSTEM AND A METHOD OF COLLECTING

AND ANALYZING A SAMPLE

Priority Claim

This application claims the benefit of U.S. Provisional Application No. 63/648,523, filed May 16, 2024, the disclosure of which is incorporated by reference herein in its entirety.

Background

[0001] The United States Centers for Disease Control and Prevention (CDC) estimates that worldwide currently two billion people may be infected with Mycobacterium tuberculosis (Mtb), with 10.6 million becoming ill with tuberculosis (TB) each year. Around 30 percent of those who become ill with TB are missed by healthcare screenings and diagnostics. Despite being preventable and treatable, TB remains a deadly disease because testing for TB is a challenge in many healthcare environments.

[0002] The current standard for testing for TB is a sputum smear test to detect Mycobacterium tuberculosis (Mtb). However, sputum is a very difficult sample type to work with. The inclusion of a lysis step in sputum tests makes sample manipulation and treatment challenging.

Additionally, producing sputum is difficult for some patients, including children and HIV patients. Sputum tests also suffer from low diagnostic sensitivity in children, HIV-infected individuals, and patients with extrapulmonary TB. Molecular assays, such as the CEPHEID GENEXPERT (available from Cepheid United States, Sunnyvale, CA, U.S.A.), can detect Mtb but are relatively expensive and difficult to operate.

[0003] The presence of Mtb can be indicated by lipoarabinomannan (LAM), which is a glycolipid found in the cell wall of Mtb. The World Health Organization (WHO) recommends urine tests for the detection of LAM to help diagnose TB. However, the testing is rendered challenging because of the difficulties associated with sample procurement and handling. Additionally, after moving through the renal system, the amount of LAM is reduced to minute quantities. Patients with HIV typically have low levels of LAM in their urine and issues with the traditional methods of sample procurement and testing can lead to high mortality outcomes for patients with HIV and TB co-morbidities. As an alternative, testing the liquid phase of exhaled air, called the exhaled breath condensate (EBC), has been explored. However, EBC sampling also suffers from shortcomings such as requiring cumbersome equipment for the collection and handling of samples.

[0004] The need exists for a simpler and more effective sample collection system with a simple- to-use procedure. Further, a need exists for an integrated sample collection system that provides samples of high purity and performs analysis. There is also a need for a more precise system to reduce human error and provide more repeatable and reliable test results.

Summary

[0005] According to an embodiment, a sample collection system includes a sample collection device including: a housing; a sample receiving element extending from the housing, the sample receiving element including an air inlet constructed to receive an exhalation sample from a single nostril; and a porous sample collection media disposed within the housing; an airflow path extending from the air inlet through the porous sample collection media; and an assay constructed to receive an eluted sample from the porous sample collection media and to detect presence of a target analyte in the eluted sample.

[0006] The sample collection system may optionally include a liquid reservoir in fluid communication with the porous sample collection media, constructed to direct liquid onto the porous sample collection media, the liquid reservoir including: a volume housing the liquid; an opening; and a cover on the opening configured to be removed, ruptured, or pierced.

[0007] The porous sample collection media may include a nonwoven filtration layer having an electrostatic charge. The nonwoven filtration layer may be hydrophobic.

[0008] A sample collection system may include a sample receiving element with a tubular body, the sample receiving element including: a first open end constructed to receive an exhalation sample from a single nostril; a second open end; and an air channel extending between the first open end and the second open end; and a sample collection device including: a sheet of porous sample collection media; a holder including a first end coupled with and constructed to support the porous sample collection media, and a second end forming a handle, wherein the sheet of porous sample collection media is removably received in the sample receiving element and arranged to receive a through-flow of the exhalation sample. [0009] The tubular body of the sample receiving element includes a wall that may have a frustoconical shape extending from the first open end toward the second open end, wherein the frustoconical shape has its smallest diameter at the first open end. The tubular body of the sample receiving element may include a slot constructed to receive the sheet of porous sample collection media.

[0010] A method of collecting and testing a sample includes flowing exhalation air from a nostril (e.g., a single nostril or both nostrils) through a porous sample collection media to form a captured sample, the porous sample collection media being disposed within a sample collection device, the sample collection device including a sample receiving element with a tubular body, the sample receiving element including: a first open end constructed to receive the exhalation air from a single nostril; a second open end; and an air channel extending between the first open end and the second open end; releasing a metered dose of liquid onto the porous sample collection media from a liquid reservoir, the metered dose of liquid eluting the captured sample onto an assay disposed within the sample collection device housing, the assay being constructed to receive an eluted sample from the porous sample collection media and to detect presence of a target analyte in the eluted sample; and observing a test result on the assay.

[0011] The observing of the test result may include observing a positive result if lipoarabinomannan (LAM) is present, or observing a negative result if LAM is absent.

[0012] A method of collecting and testing a sample may include: flowing exhalation air from a single nostril through a porous sample collection media to form a captured sample, the porous sample collection media being disposed within a sample collection device including a sample receiving element with a tubular body, the sample receiving element including: a first open end constructed to receive the exhalation air from the single nostril; a second open end; and an air channel extending between the first open end and the second open end; transferring the porous sample collection media and the captured sample to a receptacle using a holder including a first end coupled with and constructed to support the porous sample collection media, and a second end forming a handle; applying a liquid onto the porous sample collection media and the captured sample to form an eluted sample; and testing the eluted sample for presence of a target analyte using a test method. [0013] The target analyte may include a disease indicator or pathogen or both. The target analyte may include virus, bacteria, fungus, pathogen, protein, cell wall fragment, lipoarabinomannan (LAM), or a combination of two or more thereof. The target analyte may include LAM. The target analyte may include LAM and Mtb.

[0014] The assay may be constructed to detect presence of lipoarabinomannan (LAM) in a collected sample. The assay may be a lateral flow assay, a vertical flow assay, or a colorimetric indicator. The assay may have a LAM detection limit of 10 picograms/milliliters or lower.

[0015] According to an embodiment, a kit comprising the sample collection system and instructions for collecting a sample and testing the sample using an assay. The instructions may include instructions to exhale from a nostril (e.g., from a single nostril or both nostrils) through the porous sample collection media to capture a sample in the porous sample collection media, and to form a loaded porous sample collection media. The instructions may include instructions to release a metered dose of liquid onto the porous sample collection media; and observe a test result in a result display of the assay. The instructions may include instructions to transfer the loaded porous sample collection media with the captured sample to a receptacle, optionally using the holder, to apply a metered dose of liquid onto the loaded porous sample collection media to form an eluted sample; and to test the eluted sample for presence of the target analyte(s) using a test method.

[0016] The above summary is not intended to describe each embodiment or every implementation. Rather, a more complete understanding of illustrative embodiments will become apparent and appreciated by reference to the following Detailed Description of Exemplary Embodiments and claims in view of the accompanying figures of the drawing.

Brief Description of Figures

[0017] FIG. 1A is a perspective view of a sample collection system according to an embodiment.

[0018] FIG. IB is a bottom perspective view of the sample collection system of FIG. 1A.

[0019] FIG. 2A is a bottom perspective view of a top part of the sample collection system of FIG. 1A.

[0020] FIG. 2B is a top perspective view of the top part of FIG. 2A. [0021 ] FIG. 3 A is a top perspective view of a bottom part of the sample collection system of FIG. 1A.

[0022] FIG. 3B is a bottom perspective view of the bottom part of FIG. 3 A.

[0023] FIG. 4A is a top perspective view of a liquid reservoir of the sample collection system of FIG. 1A.

[0024] FIG. 4B is a bottom perspective view of the liquid reservoir of FIG. 4A.

[0025] FIG. 5A is a top view of a sample collection system according to an embodiment.

[0026] FIG. 5B is a cross-sectional side view of the system of FIG. 4A.

[0027] FIG. 5C is a perspective view of a liquid reservoir for use in the system of FIG. 4A.

[0028] FIG. 6A is a perspective view of a sample receiving element according to an embodiment.

[0029] FIG. 6B is another perspective view of the sample receiving element of FIG. 6A.

[0030] FIG. 7 is a schematic top view of a holder according to an embodiment.

[0031] FIG. 8A is a top perspective view of a sample collection device including the sample receiving element of FIG. 6A and the holder of FIG. 7.

[0032] FIG. 8B is a bottom view of the sample collection device of FIG. 8A.

[0033] FIGS. 9A and 9B are perspective views of a sample collection system including an airflow indicator according to an embodiment.

[0034] FIG. 9C is a perspective view of the airflow indicator of the sample collection system of FIG. 9A.

[0035] FIGS. 10A and 10B are perspective views of a sample collection system including an airflow indicator according to an embodiment.

[0036] FIGS. 11 A and 1 IB are perspective views of a sample collection system including an airflow indicator according to an embodiment.

[0037] FIGS. 12A and 12B are perspective views of a sample collection system including an airflow indicator according to an embodiment. [0038] FIG. 13 is a schematic partial cross-sectional view of a sample receiving element of a sample collection system including an audible airflow indicator.

[0039] FIGS. 14A-14C are schematic partial views of a sample receiving element of a sample collection system including an audible airflow indicator.

Definitions

[0040] All scientific and technical terms used herein have meanings commonly used in the art unless otherwise specified. The definitions provided herein are to facilitate understanding of certain terms used frequently herein and are not meant to limit the scope of the present disclosure.

[0041] All headings provided herein are for the convenience of the reader and should not be used to limit the meaning of any text that follows the heading, unless so specified.

[0042] Unless otherwise indicated, the terms “polymer” and “polymeric material” include, but are not limited to, organic homopolymers, copolymers, such as for example, block, graft, random and alternating copolymers, terpolymers, etc., and blends and modifications thereof.

Furthermore, unless otherwise specifically limited, the term “polymer” shall include all possible geometrical configurations of the material. These configurations include, but are not limited to, isotactic, syndiotactic, and atactic symmetries.

[0043] The term “i.e.” is used here as an abbreviation for the Latin phrase id est, and means “that is,” while “e.g.” is used as an abbreviation for the Latin phrase exempli gratia and means “for example.”

[0044] The term “about” is used here in conjunction with numeric values to include normal variations in measurements as expected by persons skilled in the art and is understood have the same meaning as “approximately” and to cover a typical margin of error, such as ±5 % of the stated value. Moreover, unless otherwise indicated, all numbers expressing quantities, and all terms expressing direction/orientation (for example, vertical, horizontal, parallel, perpendicular, etc.) in the specification and claims are to be understood as being modified in all instances by the term “about.” [0045] Terms such as “a,” “an,” and “the” are not intended to refer to only a singular entity but include the general class of which a specific example may be used for illustration.

[0046] The terms “a,” “an,” and “the” are used interchangeably with the term “at least one.” The phrases “at least one of’ and “comprises at least one of’ followed by a list refers to any one of the items in the list and any combination of two or more items in the list.

[0047] As used here, the term “or” is generally employed in its usual sense including “and/or” unless the content clearly dictates otherwise. The term “and/or” means one or all of the listed elements or a combination of any two or more of the listed elements.

[0048] The recitations of numerical ranges by endpoints include all numbers subsumed within that range (for example, 1 to 5 includes 1, 1.5, 2, 2.75, 3, 3.80, 4, 5, etc. or 10 or less includes 10, 9.4, 7.6, 5, 4.3, 2.9, 1.62, 0.3, etc.). Where a range of values is “up to” or “at least” a particular value, that value is included within the range.

[0049] As used here, “have”, “having”, “include”, “including”, “comprise”, “comprising” or the like are used in their open-ended sense, and generally mean “including, but not limited to.” It will be understood that “consisting essentially of,” “consisting of,” and the like are subsumed in “comprising” and the like. As used herein, “consisting essentially of,” as it relates to a composition, product, method or the like, means that the components of the composition, product, method or the like are limited to the enumerated components and any other components that do not materially affect the basic and novel characteristic(s) of the composition, product, method or the like.

[0050] The term “substantially” as used here has the same meaning as “significantly,” and can be understood to modify the term that follows by at least about 90 %, at least about 95 %, or at least about 98 %. The term “not substantially” as used here has the same meaning as “not significantly,” and can be understood to have the inverse meaning of “substantially,” i.e., modifying the term that follows by not more than 10 %, not more than 5 %, or not more than 2

[0051] The words “preferred” and “preferably” refer to embodiments that may afford certain benefits, under certain circumstances. However, other embodiments may also be preferred, under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful and is not intended to exclude other embodiments from the scope of the disclosure, including the claims.

[0052] Any direction referred to here, such as “front,” “back,” “top,” “bottom,” “left,” “right,” “upper,” “lower,” and other directions and orientations are described herein for clarity in reference to the figures and are not to be limiting of an actual device or system or use of the device or system. Devices or systems as described herein may be used in a number of directions and orientations.

[0053] The terms “downstream” and “upstream” refer to a relative position based on a direction of exhalation airflow through the device. For example, the upstream-most element of the device is the air inlet element, and the downstream-most element of the device is the exhalation outlet element.

[0054] The term “LAM” refers to lipoarabinomannan(s). Lipoarabinomannans are a group of compounds with a mannan backbone and branched D-arabinan and D-mannan chains. The LAM structure may be altered as it passes through the blood, renal system, and urine. The structure of LAM may vary among different strains of Mtb and can also be influenced by various factors such as the growth conditions of the bacteria and host’s immune response.

Detailed Description

[0055] The present disclosure relates to methods for testing for pathogens or other disease indicators in the respiratory tract. The present disclosure relates to sample collection devices and systems. The present disclosure relates to a device and system for collecting nasal exhalation samples, and in particular exhalation samples from a single nostril. The present disclosure further relates to a system that includes both sample collection and testing capabilities. The present disclosure further relates to methods of obtaining samples for the detection of lipoarabinomannan (LAM) from nasal exhalation samples. The present disclosure further relates to methods of testing such nasal exhalation samples for the presence of LAM in the samples. The presence of LAM can be used as an indicator of tuberculosis (TB) or the presence of Mycobacterium tuberculosis (Mtb).

[0056] The present disclosure further relates to methods of obtaining samples for the detection of one or more pathogens in nasal exhalation samples. The present disclosure further relates to methods of testing such nasal exhalation samples for the presence of Mtb or other pathogens in the samples. In most of the occurrences of tuberculosis, Mtb and LAM are available in the lungs. For 5-10 % of patients, however, TB is extra-pulmonary (established in other organs). Under certain conditions, LAM signals will be higher in the urine (possibly due to extra pulmonary TB). Thus, according to some embodiments, the nasal exhalation sample is collected along with a urine sample. The samples may be processed and tested together or separately. In some embodiments, the nasal exhalation sample is collected along with another type of sample, such as an oral or nasal swab. The nasal exhalation sample may be collected along with an oral swab sample and the samples may be processed and tested together or separately. The nasal exhalation sample may be collected along with a nasal swab sample and the samples may be processed and tested together or separately. Testing two (or more) types of samples together allows for the testing and detection of TB in patients using a single assay regardless of whether they are suffering from pulmonary or extra-pulmonary TB.

[0057] Mycobacterium tuberculosis (Mtb) is the bacterium responsible for causing tuberculosis (TB). Mtb is an acid-fast, aerobic, non-motile bacterium that primarily infects the lungs but can also affect other parts of the body may circulate and exist in many locations in a TB patient. Lipoarabinomannan (LAM) is a glycolipid found in the cell wall of Mtb. LAM is released into the bloodstream and other bodily fluids of the patient when the bacteria replicate or are killed by the immune system of the patient. Therefore, the presence of LAM in bodily fluids may serve as a biomarker for an active TB infection. However, LAM is difficult to detect in the blood due to its tendency to attach to carrier molecules.

[0058] Based on the hypothesis that LAM generated within a patient’s system may pass through to a patient’s lungs to be excreted, methods with Exhaled Breath Condensate (EBC) sampling have been studied as an alternative to the urine and blood test methods. However, EBC samples carry a risk of contamination by coming into contact with food particles, saliva, and oral microbiota, and the samples are difficult and costly to procure.

[0059] Without wishing to be bound by theory and with an aim of capturing and detecting high concentration levels of LAM with minimal contamination, it is hypothesized that due to the localization of LAM within the lungs of TB patients and the patients exhaling LAM on a regular basis, some LAM is exhaled directly, while some portion of the exhaled LAM is caught and concentrated in the mucosal layers lining the epithelium of the respiratory tract. Thus, nasal exhalation samples from patients result in a higher concentration of the analytes of interest (for example, LAM) in the samples, and may be used for the indication of the presence of Mtb and/or active TB infection. Devices, systems, and methods for such sampling and analysis are provided in the present disclosure. These devices, systems, and methods may also be used for sampling and analysis of any other analytes (e.g., disease indicators and/or pathogens) that may be present in the respiratory tract. Such other analytes may include virus, bacteria, fungus, pathogen, or other analytes. In embodiments where the target analyte is a virus, it may be enveloped or nonenveloped. The virus may be a coronavirus, rhinovirus, norovirus, influenza virus, adenovirus, adeno-associated virus, varicella-zoster virus, herpesvirus, retrovirus, papillomavirus, enterovirus, arenavirus, or another type of virus. In some embodiments, the virus is present in a respiratory tract sample. In embodiments wherein the target analyte is a coronavirus, it may be SARS-CoV-2 virus (e g., COVID-19). In some embodiments, the target analyte is bacteria. The bacteria may be, for example, Mycobacterium tuberculosis (Mtb), Streptococcus pneumoniae, Mycoplasma pneumoniae, Haemophilus influenzae, Chlamydophila pneumoniae, Coxiella burnetiid, Moraxella catarrhalis, Histoplasma capsulatum, or Legionella pneumophila. In some embodiments, the target analyte is a fungus. The fungus may be, for example, Cryptococcus neoformans, Aspergillus, Pneumocystis, or endemic fungi. The target analyte may be a disease indicator, such as a protein or cell wall fragment.

[0060] In some embodiments, the methods of the present disclosure include sampling and analysis of respiratory tract samples for the presence of LAM and Mtb. In some embodiments, the devices and systems of the present disclosure include sampling and analysis of respiratory tract samples for the presence of LAM and Mtb.

[0061] It has also been discovered that lower amounts of foreign proteins or enzymes (such as cleaning chemicals) are present in the nasal exhalation samples, leading to lesser chances of contamination. Additionally, as described in the present disclosure, it has also been discovered that the concentration of analytes (for example, LAM or other analyte or pathogen) is further increased in nasal exhalation samples by closing one nostril to breathe with the opposite open nostril to create adequate pressure and back-pressure. The pressure and backpressure create turbulence that dislodges some of the mucosal fdm along the surface of the epithelium of both the upper and the lower respiratory tracts. The air flow from the lungs may be constricted in the passages of the nasal cavity, causing increased velocity and turbulent flow, which in turn causes material to be dislodged from the surfaces of the nasal cavity.

[0062] Another drawback of using EBC to detect LAM is that EBC contains predominately oral fluids, while the samples collected by the devices and methods of the present disclosure substantially avoid the presence of oral fluids. This may allow concentration of the signal due to the increased analyte availability in the sample that includes dislodged mucosal particulate from the respiratory tract’s epithelial cells, and the elution into a relatively small volume of liquid (e.g., buffer) to elute the analyte.

[0063] The term “respiratory tract sample” is used here to refer to a sample obtained by exhalation through the nose. A respiratory tract sample includes material (e.g., mucus, cell fragments, and potentially disease markers) present in the respiratory tract. A respiratory tract sample may include material from the entire respiratory tract, including the nasal cavity. A respiratory tract sample is distinguished from an oral exhalation sample that mainly includes exhalation air and some moisture from the lungs but minimal or no material from the respiratory tract. Oral exhalation samples typically do not include material from the nasal cavity as the orally exhaled air does not pass through the nasal cavity.

[0064] In order to obtain a respiratory tract sample, a user may exhale through one or both nostrils. The user may close one nostril and exhale through a single nostril. Closing one nostril increases the flow rate through the open nostril and may increase the amount of material from the respiratory tract in the sample. Typical exhalation flow rates for adults may range from about 30 L/min (at rest) to about 85 L/min (at high effort). Forcefully exhaling through a single nostril may produce a flow rate in the upper range, e.g., from about 50 L/min to about 90 L/min. The sample collection devices of the present disclosure are constructed to receive nasal exhalation samples at flow rates of 30 L/min or greater, 40 L/min or greater, 50 L/min or greater, 60 L/min or greater, or 70 L/min or greater. The sample collection devices of the present disclosure are constructed to receive nasal exhalation samples at flow rates of 120 L/min or lower, 110 L/min or lower, 100 L/min or lower, 90 L/min or lower, or 80 L/min or lower.

[0065] In some embodiments, the nasal exhalation sample is collected along with a urine sample. In some embodiments, the nasal exhalation sample is collected along with a nasal swab sample. In some embodiments, the nasal exhalation sample is collected along with an oral swab sample. The samples may be processed and tested together or separately.

[0066] The present disclosure provides a sample collection device constructed for collecting a nasal exhalation sample. The sample collection device includes porous sample collection media constructed to capture the target analyte from nasal exhalation. The sample collection media may be housed in a housing or supported by a holder. The housing or holder may be configured to obtain a sample via nasal exhalation. The sample collection device may optionally include an exhalation piece (a sample receiving element) that helps guide a nasal exhalation sample from a nostril or nostrils onto the sample collection media. The sample collection device may include a tubular sample receiving element with a first open end and a second open end with an air channel extending between the two ends. According to an embodiment, the first end is constructed to receive an exhalation sample from a single nostril. For example, the first end may have a shape that is suitable for obtaining an exhalation sample from a single nostril. The first end may have a size that is suitable for obtaining an exhalation sample from a single nostril. The first end may be made of a material that is suitable for obtaining an exhalation sample from a single nostril. The tubular body of the sample receiving element may have a frustoconical shape extending from the first open end toward the second open end, where the frustoconical shape has its smallest diameter at the first open end (e.g., adjacent the air inlet). Such a shape may be particularly suitable for accommodating nostrils of different sizes and shapes, thus enabling collection of a nasal exhalation sample from most users. The system further includes a sample collection device with a porous sample collection media. The sample collection media may be housed or held in a device that facilitates collection of the sample without contamination of the sample collection media with contaminants from the environment or, for example, the user’s fingers. For example, the sample collection media may be disposed within the device housing and along an airflow path defined by the device housing. The device housing may be coupled with the sample receiving element. The device housing may include an air inlet to receive the exhalation sample from the sample receiving element. The porous sample collection media is constructed to capture the target analytes, such as LAM, carried in an exhalation airflow.

[0067] The target analyte captured by the porous sample collection media may be a disease indicator and/or pathogen. The target analyte captured by the porous sample collection media may be virus, bacteria, fungus, pathogen, or other analyte (e.g., protein or cell wall fragment). The target analyte captured by the porous sample collection media may be LAM. The target analytes captured by the porous sample collection media may be LAM and Mtb.

[0068] The system may further include a sample testing assay. The system of the present disclosure may include a sample testing assay to test for the presence of a disease indicator and/or pathogen in the sample. The system may include a sample testing assay to test for the presence of virus, bacteria, fungus, pathogen, or other analyte (e.g., protein or cell wall fragment) in the sample. The system may include a sample testing assay to test for the presence of LAM in the sample. The system may include a sample testing assay to test for the presence of LAM and Mtb in the sample.

[0069] The system may further include a liquid, such as a metered dose of liquid, to facilitate the elution of the sample from the sample collection media and the testing of the sample with the assay. The metered dose of liquid may be provided in a liquid reservoir provided within the housing or as part of a kit. The metered dose of liquid may be passed through the porous sample collection media and may carry away the analytes that may be bound to the porous sample collection media, forming an eluent. The analyte bound to the porous sample collection media may be a disease indicator and/or pathogen. The analyte bound to the porous sample collection media may be virus, bacteria, fungus, pathogen, or other analyte (e.g., protein or cell wall fragment). The analyte bound to the porous sample collection media may be LAM. The analyte bound to the porous sample collection media may be Mtb. The analyte bound to the porous sample collection media may include both LAM and Mtb. The eluent may include eluted disease indicator and/or pathogen. The eluent may include eluted virus, bacteria, fungus, pathogen, or other analyte (e.g., protein or cell wall fragment). The eluent may include eluted LAM. The eluent may include eluted Mtb. The eluent may include eluted LAM and Mtb. The eluent may further flow onto the assay and be analyzed using the assay. The eluent may be analyzed using a separate test method.

[0070] In some embodiments, the present disclosure relates to a sample collection device including a housing extending from an air inlet to an air outlet. The housing defines an airflow path from the air inlet to the air outlet. The air inlet is configured to receive an exhalation airflow. The exhalation airflow may be nasal exhalation air. A porous sample collection media is disposed within the housing and along the airflow path. According to an embodiment, the sample collection device includes a nosepiece. In the present disclosure, the nosepiece is sometimes referred to as a sample receiving element. In some embodiments, the sample receiving element includes both a nosepiece and a mouthpiece. That is, the sample receiving element may include structures that facilitate breathing or exhaling into an air inlet either via a nostril or the mouth. The sample receiving element may be aligned with the inlet opening of the device housing. The sample receiving element may help a user direct exhalation airflow onto the porous sample collection media. The sample receiving element may be integral with the device housing or may be a separate part. In some embodiments, flowing exhalation air includes blowing into the sample receiving element on the sample collection device.

[0071 ] In its simplest form, the sample collection device of the present disclosure includes porous sample collection media, and some type of a holder or housing constructed to support the porous sample collection media such that the sample collection media is arranged to receive a through-flow of exhalation sample from a user’s nostril(s). The user may hold the sample collection device against his or her nostril and blow through the nostril into or onto the sample collection device and through the porous sample collection media such that a nasal exhalation sample is deposited onto the sample collection media. The sample collection device may include a sample receiving element. The sample receiving element is configured to receive a nasal exhalation sample and to direct the sample into the inlet or the housing or directly onto the porous sample collection media. The holder or housing may optionally include further features, such as elements that facilitate the user holding the device and obtaining a sample, or features that facilitate eluting and testing of the sample. Such features are further discussed in detail below.

[0072] Reference is made in the following to the various views of the drawings. However, it is understood that not all combinations of the features are shown, and other combinations may exist.

Sample Receiving Element

[0073] The sample receiving element may be provided as a separate part, as shown in FIGS. 6A and 6B, or may formed integrally with the sample collection device housing, as shown in FIGS. 1A-12A. [0074] Generally, the sample receiving element 10 has a first end 11, a second end 12, and an air channel 15 extending from the first end to the second end. The first end and second ends 11, 12 are both open ends. The sample collection devices shown in FIGS. 1A, 9A, 10A, 11A, and 12A include an integral sample receiving element 10’, 1010, 2010, 3010, 4010. The sample receiving element 10, 10’, 1010, 2010, 3010, 4010 may have any suitable size and shape that facilitates obtaining a nasal exhalation sample from a single nostril. Such a sample receiving element may be referred to as a “nose cone.” As noted above, it is hypothesized that by closing one nostril and forcefully exhaling through the open nostril allows for the collection of a sample that includes some of the mucosal film along the surface of the epithelium of both the upper and the lower respiratory tracts and may provide an adequate sample from which the presence of analytes (for example, LAM) may be tested. To facilitate obtaining such as sample, the sample receiving element may have a shape and size that allows the first end of the sample receiving element to be at least partially inserted into the nostril. The shape and size of the first end of the sample receiving element may be such that a sealing fit with the nostril of most users may be obtained. While the sample receiving element 10 may generally have a tubular shape (e.g., have a wall 13 circumscribing a hollow interior 14 with two opposing open ends), the sample receiving element 10 may have a cross-sectional diameter D10 that is smaller at the first end 11 and larger toward the second end 12. The first end 11 may have a frustoconical shape extending from the first open end toward the second open end, where the frustoconical shape has its smallest diameter at the first end 11. The smallest diameter of the sample receiving element 10 may be about 6 mm to about 10 mm. The largest diameter of the sample receiving element 10 may be about 10 mm to about 17 mm or about 12 mm to about 15 mm. In some embodiments, the frustoconical shape continues along the entire length of the sample receiving element 10 from the first end 11 to the second end 12. That is, the cross-sectional diameter of the sample receiving element may increase from the first end 11 to the second end 12. The frustoconical shape provides a smooth change in diameter or width from the smallest diameter or width at the open end to the larger diameter or width in the direction of the second open end. This shape allows for an efficient collection of nasal exhalation samples from nostrils of varying shapes and sizes. In some embodiments, the sample receiving element may have a substantially frustoconical shape for insertion into the nostril of a patient to provide a seal between the sample receiving element and the nostril of the patient. In some embodiments, the frustoconical shaped sample receiving element may not provide a sealing fit with the nostril. In some embodiments, the frustoconical shape of the sample receiving element provides for the formation of at least a partial seal between the underside of the nose of the patient and the nosepiece. The cross-sectional shape of the sample receiving element (in a cross-section taken perpendicular to the air channel) may be round or obround. Here, the shape and size of the sample receiving element refers to the outside shape and size, e.g., the outside dimensions of the sample receiving element 10. The shape and size of the air channel 15 inside the sample receiving element may follow the overall shape and size of the sample receiving element, or may be different from the overall shape and size (e g., have the same inner diameter throughout the length of the air channel).

[0075] The sample receiving element may be made of a material that allows the sample receiving element to be shaped to the user’s nostril. For example, the sample receiving element may be made of a flexible polymer, such as rubber or silicone. The shape, size, and material of the sample receiving element allow for the delivery of the nasal exhalation sample from the nasal cavity of the patient to the sample receiving element of the sample collection device. The nasal exhalation sample is delivered from the nasal cavity of the patient to the sample receiving element of the sample collection device through the inner air channel of the frustoconical shaped sample receiving element extending from the first open end to the second open end.

[0076] In some embodiments, the second open end 12 of the sample receiving element 10 may be constructed for receiving a breath sample from a user’s mouth. In some embodiments, the user may exhale from the mouth into the second end 12 of the sample collection device 10. That is, the user may exhale through either the first end 11 (the nasal sampling end) or the second end 12 (the mouth sampling end).

[0077] In some embodiments, as shown in FIG. 6B, the sample receiving element 10 includes a slot 16 or a track for inserting sample collection media such that the sample collection media occludes the airflow channel. The sample receiving element 10 may include a slot 16 or a track for inserting sample collection media held by a holder (see FIGS. 8A and 8B). The sample receiving element 10 may include an indicator hole 17 through which a user can see whether the holder 160 is fully inserted into and seated in the sample receiving element 10. [0078] The sample receiving element 10 may be integral with the sample collection device. That is, the sample receiving element 10 may be formed by an extension in the housing 100 of the sample collection device as shown in FIGS. 1A, 2A, and 2B.

Sample Collection Media

[0079] The sample collection device includes porous sample collection media constructed to capture the target analyte from an exhaled sample, such as via nasal exhalation. The sample collection media may be disposed within a housing or alternatively held in or supported by a holder. According to an embodiment, the porous sample collection media is disposed in an airflow path of a sample receiving element, extending from the air inlet through the porous sample collection media.

[0080] The user may exhale from a nostril (or alternatively both nostrils) into the sample receiving element, from which the sample is directed into the sample collection device along the airflow channel of the sample receiving element and the airflow path of the sample collection device. The exhalation sample loads the porous sample collection media to form a loaded porous sample collection media. In some embodiments, the user may exhale from the mouth into the sample receiving element. For example, the user may exhale through the first end of the sample receiving element (configured as a nosepiece), or through the second end of the sample receiving element (configured as a mouthpiece). The housing may be constructed such that by exhaling through the single opening, air inlet, nosepiece, or mouthpiece, the exhalation airflow passes through the porous sample collection media. The porous sample collection media is constructed to capture analytes (for example, LAM), from the exhalation airflow.

[0081] The porous sample collection media is suitable for exhalation through the media. That is, the porous sample collection media has sufficient porosity to allow exhalation through the media. As used here, the term “porosity” refers to a ratio of open space in the media to the amount of volume taken by the media material itself. A media with high porosity has more open space and, therefore, allows higher flow with a lower pressure drop.

[0082] In some embodiments, the porous sample collection media at least partially occludes the airflow path within the sample collection device. The porous sample collection media may completely occlude the airflow path. [0083] Exhalation airflow may pass through the thickness of the porous sample collection media. Components, such as target analytes, may become caught in the sample collection media, thus forming loaded sample collection media (or simply “loaded media”). The porous sample collection media may have a major plane that is orthogonal to the direction of the exhalation airflow passing through the thickness of the porous sample collection media. In some embodiments, the porous sample collection media is arranged (e.g., folded, pleated, or otherwise shaped) such that it is at an angle relative to the direction of the exhalation airflow passing through the thickness of the porous sample collection media.

[0084] In some embodiments, the porous sample collection media may include a nonwoven filtration layer. In some embodiments, the porous sample collection media has an electrostatic charge. In some embodiments, the porous sample collection media may include a nonwoven filtration layer having an electrostatic charge. The electrostatic charge may enable capturing analytes (for example, LAM) from an exhalation airflow. In some embodiments, the porous sample collection media is hydrophobic. In some embodiments, the nonwoven filtration layer may be a hydrophobic nonwoven filtration layer. The nonwoven filtration layer may be a hydrophobic nonwoven filtration layer carrying an electrostatic charge configured to capture analytes (for example, LAM) from an exhalation airflow. In some embodiments, the porous sample collection media is hydrophilic. In some embodiments, nonwoven filtration layer may be a hydrophilic nonwoven filtration layer. The nonwoven filtration layer may be a hydrophilic nonwoven filtration layer carrying an electrostatic charge configured to capture analytes (for example, LAM) from an exhalation airflow. The term “hydrophobic” refers to a material having a water contact angle of 90 degrees or greater, or from about 90 degrees to about 170 degrees, or from about 100 degrees to about 150 degrees. The term “hydrophilic” refers to a material having a water contact angle of less than 90 degrees. Water contact angle is measured using ASTM D5727-1997 Standard test method for surface wettability and absorbency of sheeted material using an automated contact angle tester.

[0085] The porous sample collection media may be formed of any suitable material that is capable of capturing analytes (for example, virus, bacteria, fungus, pathogen, LAM, or other analyte (e.g., protein or cell wall fragment), or a combination thereof) from exhalation airflow and releasing the captured analytes upon being contacted with an eluent, such as a saline solution. The porous sample collection media may be formed of polymeric material. The porous sample collection media may be formed of a polyolefin. Examples of suitable polyolefins include polypropylene, polylactic acid, and the like, and a combination thereof. In one embodiment the porous sample collection media is formed of polypropylene. In one embodiment the porous sample collection media is formed of polylactic acid. One illustrative porous sample collection media is commercially available from 3M Company (St. Paul, MN, U.S.A.) under the trade designation FILTRETE Smart MPR 1900 Premium Allergen, Bacteria & Virus Air Filter Merv 13.

[0086] The porous sample collection media may have a thickness (orthogonal to the major plane) of 200 pm or greater, or 250 pm or greater. The porous sample collection media may have a thickness of 750 pm or less, or 1000 pm or less. The porous sample collection media may have a thickness of in a range from 200 pm to 1000 pm, or from 250 pm to 750 pm. The porous sample collection media may have major plane surface area (of one side) of 1 cm² or greater, or 2 cm² or greater. The porous sample collection media may have major plane surface area of 3 cm² or less, or 4 cm² or less. The porous sample collection media may have major plane surface area in a range from 1 cm² to 4 cm², or 2 cm² to 3 cm². The porous sample collection media defines a surface area, and the volume of the liquid reservoir divided by the surface area is in a range from 10 pm/cm² to 400 pm/cm², or from 10 pm/cm²to 250 pm/cm².

Housing

[0087] The sample collection device may include a housing constructed to support the porous sample collection media such that the sample collection media is arranged to receive a through- flow of exhalation sample from the sample receiving element.

[0088] The shape, size, and form of the housing is not particularly limited. Various types of sample collection device housings suitable for use in the system of the present disclosure are described, for example, in PCT/US2021/034327 fded on 26 May 2021; PCT/US2021/041485 fded on 13 Jul 2021; PCT/IB2022/059989 filed on 18 Oct 2021; PCT/US2022/012392 filed on 14 Jan 2022; PCT/US2022/014388 filed on 28 Jan 2022; PCT/IB2022/051250 filed on 11 Feb 2022; PCT/IB2022/051251 filed on 11 Feb 2022; PCT/IB2022/051252 filed on 11 Feb 2022; PCT/US2022/019399 filed on 8 Mar 2022; PCT/IB2022/054633 filed on 18 May 2022; PCT/IB2022/054683 filed on 19 May 2022; PCT/IB2022/054852 filed on 19 May 2022; PCT/IB2022/056708 filed on 20 July 2022; PCT/IB2022/056714 filed on 20 July 2022; each of which is incorporated herein by reference.

[0089] Referring now to the drawings, it should be noted that, as used here, like reference numbers refer to like elements throughout the various views in the drawings. Further, in some cases a similar part may be designated using a number and a prime (e.g., 10 and 10’). It should be understood that, unless otherwise noted or is otherwise clear from the context, the description of the part with (or without) the prime applies equally to the part without (or with) the prime.

[0090] An exemplary embodiment of an integrated sample collection system 1 that includes a sample collection device and assay in shown in FIGS. 1A, IB, 5A, and 5B. The sample collection device has a housing 100 that houses both the porous sample collection media 120 and an assay 300. The housing 100 may further house a liquid reservoir 200. A sample receiving element 10’ is integral with the sample collection device housing 100. The housing 100 defines an airflow path 110 and houses the porous sample collection media 120. In some embodiments, the housing 100 includes a first end 101 and an opposing second end 102. The sample receiving element 10’ includes an air channel 15’, which may form an air inlet 131 adjacent the first end 101 and a part of the airflow path 110 which extends from the air inlet 131 through the porous sample collection media 120.

[0091] Another example of a housing 100’ that may be modified to be used with the sample receiving element 10 of FIG. 6A is shown in FIGS. 5A and 5B. The housing 100’ defines an airflow path 110’ and houses the porous sample collection media 120. In some embodiments, the housing 100’ includes a first end 101’ and an opposing second end 102’. An air inlet 131 is formed adj acent the first end 101’. The air inlet 131 defines a part of airflow path 110’ which extends from the air inlet 131 through the porous sample collection media 120.

[0092] The housing 100 may be formed in one piece or may be formed of two or more pieces or parts that together form the housing. In particular in embodiments where the sample collection system 1 includes an assay 300, the housing 100 may be formed of two parts 111, 111. Referring now to FIGS. 1 A-3B, , the housing 100 includes a first part 111 coupled with a second part 112. The first part 111 may be a top part and the second part 112 may be a bottom part. The first and second parts 111, 112 may be sandwiched together, capturing within the housing 100 the sample collection media 120, a liquid reservoir 200, and an assay 300. The sample receiving element 10 and air inlet 130 may be formed in the first part 1 11 . An airflow path 110 may be formed through the first part 111 and the sample collection media 120. The airflow path 110 may further extend through outlet(s) 134 in the second part 112, as shown in FIGS. IB, 3A, and 3B. The porous sample collection media 120 may be captured between the first and second parts 111, 112 and arranged to occlude the airflow path 110. An assay 300 may also be captured between the first and second parts 111, 112. The first part 111 may include a result viewing window 170 through which the assay result area 370 may be viewed. The first part 111 and second part 112 may be connected by a snap fit closure or may be sealed or adhered together. The first and second parts 111, 112 may include corresponding coupling features 154 (first part 111, FIG. 2 A) and 155 (second part 112, FIG. 3 A).

[0093] The housing may be formed of a rigid material, such as plastic or a paper-based material such as cardboard or cardstock. In some embodiments, the housing is made of plastic. The housing may be made of a material that does not absorb any of the liquid or eluent. For example, the housing may be made of a hydrophobic material. In some embodiments, at least a portion of the housing is transparent. For example, the housing may include transparent material in an area of a result display of an assay.

[0094] In some embodiments, the housing includes or houses an assay, as further discussed below. The housing may also include or house a liquid reservoir, as also further discussed below.

[0095] In some embodiments, the housing includes an indicator that indicates whether sufficient airflow has been applied to obtain a sample. An airflow indicator may be particularly useful in a sample collection device that is configured for obtaining a sample by nasal exhalation. When the sample is obtained by nasal exhalation, there may be a concern that users do not apply sufficient air flow to provide a reliable sample, which could result in false negative results.

[0096] The airflow indicator may be configured to provide a visual, audible, or tactile indication of sufficient airflow, or a combination of two or more thereof. For example, the airflow indicator may indicate sufficient airflow by the appearance or movement of an element (e.g., a marker, an extension, or a tab) and/or appearance of a color that can be tactilely or visually observed, or by a sound (e.g., a whistle) that can be audibly observed. In some embodiments, the indicator includes a movable part that is configured to move from a first position to an indicating position once the airflow exceeds a predetermined threshold. In some embodiments, the indicator may include an element connected to the movable part that provides resistance by friction or spring force. Tn some embodiments, the movable part may move gradually across indicators that, once sufficient amount of air has passed through the housing, the movable part indicates a positive indicator. Examples of airflow indicators are shown in FIGS. 9A-14C. Such indicators may be combined with any of the housing types discussed above.

[0097] Referring now to FIGS. 9A-10B, sample collection systems with a sample collection device housing 1100, 2100 are shown. The type of housing is not particularly limited as the airflow indicator may be applied to any of the housing types discussed above. The housing 1100, 2100 may be similar to the housing discussed above with regard to FIGS. 1A-3B, including a first end 101 and an opposing second end 102, a sample receiving element 1010, 2010 an air inlet 131, porous sample collection media, an airflow path 1110, 2110 that extends from the air inlet 131 through the porous sample collection media, an assay 300, and a test result display window 1170, 2170. The housing 1100, 2100 may further house a liquid reservoir that in the examples shown is a blister 1200, 2200 containing a metered dose of liquid. The blister 1200, 2200 is configured to be ruptured or punctured when pressure is applied to the blister 1200, 2200 to release the metered dose of liquid.

[0098] The airflow applied to the air inlet 131 creates an increased pressure inside the housing 1100, 2100. The housing 1100, 2100 includes an airflow indicator 1400 with a spring mechanism that provides resistance to the airflow. Once the force of the airflow exceeds the threshold spring force, the airflow indicator 1400 extends out of the housing 1100, 2100, providing a positive indicator of sufficient airflow. The airflow indicator 1400 may be disposed at the second end 102 of the housing 1100, 2100. The housing 1100, 2100 includes an aperture 1120, 2120 through which the airflow indicator 1400 extends. The airflow indicator 1400 includes a tab 1410 and one or more springs 1420. The one or more springs 1420 create resistance to the airflow. The housing 1100, 2100 may include a corresponding groove or track in which the airflow indicator 1400 is disposed. The one or more springs 1420 may be tuned such that the tab 1410 slides out of the housing 1100, 2100 only with sufficient airflow. The tab 1410 provides a visual and a tactile indicator of the sufficient airflow. The tab 1410 may further include a color indicator 1412 to provide a visual indicator of the sufficient airflow. [0099] In another example, also applicable to any of the housing types discussed above, an airflow indicator 3400, 4400 with a marker 3410, 4410 is configured to slide along an indicator track 3430, 4430 as shown in FIGS. 11A-12B. In the first embodiment (FIGS. 11A and 1 IB), the indicator 3400 includes a color indicator. In the second embodiment, the indicator 4400 includes a character indicator, for example including characters such as (minus sign) and “+” (plus sign). The color indicator and the character indicator include a first portion 3431, 4431 that indicates insufficient airflow and a second portion 3432, 4432 that indicates sufficient airflow. The marker 3410, 4410 moves along the indicator track 3430, 4430, and moves from a first position within the first portion 3433, 4433 to a second position within the second portion 3434, 4434 when sufficient amount of airflow has been applied. The marker 3410, 4410 and indicator track 3430, 4430 may also constructed such that a certain amount of air pressure is needed to move the marker 3410,4410 in the indicator track 3430, 4430. This way, the marker 3410, 4410 may indicate both sufficient amount and sufficient pressure of airflow. A user may also exhale through the device multiple times if a single exhalation does not move the marker 3410, 4410 all the way from the first portion 3431, 4431 to the second portion 3432, 4432.

[0100] Examples of audible airflow indicators 450 are shown in FIGS. 13 and 14A-14C. An audible airflow indicator 450 may be positioned anywhere along the airflow channel 15 and airflow path 110. In some embodiments, the audible airflow indicator 450 may be disposed in the interior 14 of the sample receiving element 10 (e.g., nose cone), as shown schematically in FIG. 13. The audible airflow indicator 450 may include an element that produces a sound when airflow is applied to the element. For example, the audible airflow indicator 450 may include an element that forms a whistle or a vibrating membrane or diaphragm. An example of a whistletype airflow indicator 5450 is shown in FIGS. 14A-14C. The airflow indicator 5450 includes a slot 5452 that creates a whistling sound when airflow is blown past it. The slot 5452 may be configured such that it emits the whistling sound only when it is exposed to a sufficient velocity of airflow. The airflow indicator 5450 may be provided as an insert 5454 disposed in the interior 5014 of the sample receiving element 5010. Alternatively, an airflow indicator 450 may be provided as an integral part of the housing 100, e.g., the sample receiving element 10. Holder

[0101] In some embodiments, the sample collection media 120 is held by a holder 160, shown in FIG. 7. A holder 160 may be distinguished from a housing in that it may only partially encapsulate the sample collection media and/or it may lack some of the other components, such as features that facilitate the presence of an assay or liquid reservoir. A holder may provide support for the sample collection media, a means for coupling the sample receiving element with the sample collection media, and/or features that facilitate handling and holding the sample collection media.

[0102] In some embodiments, the holder 160 provides support for the sample collection media 120’. The holder may include a frame 162 or other support structures that hold the sample collection media 120’ in place. In some embodiments, the holder 160 includes a handle 270 for holding and/or handling the sample collection media. The handle 270 may extend from a first end 271 at the sample collection media 120’ to a second end 272. The frame 162 or other support structure may at least partially surround the sample collection media 120’ and the handle 270.

[0103] The sample receiving element 10 may include a slot 16 for sliding the sample collection media 120’ (held by the holder 160) into, thus placing the sample collection media 120’ in the air channel 15 of the sample receiving element 10, as shown in FIGS. 8A and 8B.

Sample Collection System

[0104] In some embodiments, the sample collection device is part of a sample collection system. The sample collection system of the present disclosure includes a sample receiving element as described above, constructed to receive an exhalation sample from one or two nostrils, or for example a single nostril. The sample collection system further includes a sample collection device. The sample collection device includes the porous sample collection media. The sample collection device may include a housing or a holder. The housing or holder is constructed to support the porous sample collection media such that the sample collection media is arranged to receive a through-flow of exhalation sample from the sample receiving element. The sample collection system may further include an assay, as discussed below. The sample collection system may further include a liquid reservoir, optionally containing a metered dose of liquid, as discussed below. [0105] The sample collection system 1 may include an assay 300 housed in the housing 100 as discussed above with regard to FIGS. 1A, 2A, 5A, and 5B. The housing 100 may also be constructed to facilitate housing a liquid reservoir 200 (e.g., a capsule) including a metered dose of a liquid 210. Exemplary liquid reservoirs 200 are shown in FIGS. 4A, 4B, and 5C. The liquid reservoir 200 has an interior volume V200. The liquid reservoir 200 may optionally include features that facilitate placement of the liquid reservoir 200 inside the housing 100 and may help keep the liquid reservoir 200 in place during use. The liquid reservoir 200 may be sealed by a removable closure including a removable tab 250 (see FIGS. 1A, IB, and 5A-5C). After loading the sample collection media 120 with a nasal exhalation sample, the user may release the metered dose of liquid 210 from the liquid reservoir 200 by pulling on the removable tab 250. The metered dose of liquid 210 elutes the sample and flows or wicks the sample onto the assay 300. The result of the assay may be observed through a window 170 on the housing 100.

[0106] Alternatively, the sample collection device may be used with a separate liquid reservoir. Such separate liquid reservoirs may include a dropper or a liquid capsule with a membrane or seal that is removable, breakable, or pierceable.

Assay

[0107] In some embodiments, the sample collection system 1 includes the sample collection device described above and an assay 300 constructed to receive an eluted sample from the porous sample collection media and to detect the presence of a target analyte in the eluted sample. The assay may be constructed to detect presence of a virus, bacteria, fungus, pathogen, or other analyte (e.g., protein or cell wall fragment) in the sample. In some embodiments, the assay is constructed to detect presence of LAM in a collected sample. The assay may be constructed to receive a metered dose of liquid (for example, the eluent) from the porous sample collection media. The assay may further be constructed to analyze the target analyte, including virus, bacteria, fungus, pathogen, LAM, or other analyte (e.g., protein or cell wall fragment).

[0108] The assay may be integrated with the sample collection device. The assay may form a unitary element with the housing of the sample collection device. The user may exhale from the nose or mouth into the sample receiving element and load the porous sample collection media with a sample of the exhalation airflow to form a loaded porous sample collection media. The user may then release a metered dose of liquid from a liquid reservoir to apply a liquid to the loaded porous sample collection media and to elute the captured sample onto the assay. The user may test the eluent for the presence of a target analyte using the assay. The user may test the eluent for the presence of a virus, bacteria, fungus, pathogen, or other analyte (e.g., protein or cell wall fragment) using the assay. The user may test the eluent for the presence of LAM using the assay. The testing may take place with the loaded porous sample collection media in place in the sample collection system.

[0109] The assay included in the sample collection system may be any suitable assay. The assay may be configured to determine the presence or absence of a target analyte (for example, virus, bacteria, fungus, pathogen, LAM, or other analyte (e.g., protein or cell wall fragment)), in the collected sample. In some embodiments, the assay is a lateral flow assay (“LFA”), a vertical flow assay (“VFA”), or a colorimetric indicator. LFAs and VFAs are generally paper-based platforms for the detection and quantification of analytes in complex mixtures, including biological samples such as saliva, urine, etc. LFAs and VFAs are typically easy to use and can be used both by professionals in a health care setting or laboratory as well as by lay persons at home. Typically, a liquid sample is placed on the assay in a sample receiving region and is wicked by capillary flow along the device to a test region. LFAs and VFAs are typically based on antigens or antibodies that are immobilized in the test region and that selectively react with the analyte of interest. The result is typically displayed within 5 to 30 minutes. LFAs and VFAs can be tailored for the testing of analytes (for example, LAM). According to an embodiment, the assay used in the sample collection system of the present disclosure is constructed for the detection of an analyte (for example, virus, bacteria, fungus, pathogen, LAM, or other analyte (e.g., protein or cell wall fragment)). According to an embodiment, the assay used in the sample collection system of the present disclosure is constructed for the detection of an analyte, that may be present in the exhalation air flow of a subject. According to an embodiment, the assay used in the sample collection system of the present disclosure is constructed for the detection of LAM, that may be present in the exhalation air flow of a subject.

[0110] The assay may be capable of detecting LAM at a picogram level. For example, some assays may be capable of detecting as low as 10 pg of LAM. The LAM may be eluted with a metered dose of liquid, e.g., with 100 pL to 1000 pL, or with about 250 pL of liquid (e.g., buffer). The concentration of LAM in the eluted sample may be, for example, about 10 pg/mL to about 1 pg/mL. The assay may be capable of detecting LAM at these levels. That is, the assay may have a limit of detection (“LOD”) for LAM of about 10 pg/mL.

[0111] Examples of colorimetric indicators include LFA colorimetric readers utilizing image sensors, such as a charge-coupled device (CCD) or complementary metal oxide semiconductor (CMOS). Such devices are useful, at least in part, due to their simple structure and small size. When the device is used, the LFA develops a test line, which is an aggregate of labeled particles, antigens, and antibodies. An image sensor-based LFA reader acquires an image of the test line analyzes the pixel intensity of the test line, which changes according to the concentration of the target analyte.

[0112] According to an embodiment, the sample collection system forms a singular self- contained unit. Providing a self-contained unit allows for convenient shipping and transportation of the sample collection system and for disposal after use. The self-contained unit may have a compact size and may be conveniently carried in a pocket or purse. The self-contained unit may be safely disposed of after use among ordinary waste disposal.

[0113] The assay may be a separate element from the sample collection device. The assay may be configured to attach to the sample collection device. The sample collection device may include a receptacle for receiving at least a portion of the assay. The sample collection device may include a receptacle for receiving the entire assay. The assay may be a replacement element with the sample collection device.

Liquid Reservoir and Metered Dose of Liquid

[0114] In some embodiments, the sample collection system may include a liquid reservoir. The liquid reservoir is discussed above with regard to FIGS. 4A, 4B, and 5C. The liquid reservoir may be disposed within the housing. The liquid reservoir may contain a metered dose of a liquid. The liquid reservoir may be in fluid communication with the porous sample collection media, and constructed to direct a liquid onto the porous sample collection media. In some embodiments, a liquid flow channel may extend between the liquid reservoir and the porous sample collection media. The liquid reservoir may be formed by the housing itself or may be provided as a liquid capsule disposed within the housing. The liquid reservoir may be constructed to release a metered dose of liquid onto the porous sample collection media. The liquid reservoir may include a volume housing the liquid, an opening, and a cover on the opening configured to be removed, ruptured, or pierced. In some embodiments, optionally the liquid reservoir may be immediately adjacent to the air inlet. The liquid reservoir (for example, liquid capsule) may be prepared from any suitable material, such as polyethylene (PE), polypropylene (PP), polyester terephthalate (PET), or the like. The liquid reservoir may house any desired amount of liquid, such as from 50 microliters to 1000 microliters.

[0115] In some embodiments, the liquid reservoir may overlap the porous sample collection media. Alternatively, the liquid reservoir may be in fluid communication with (for example, adjacent or immediately adjacent) the porous sample collection media such that a liquid from the liquid reservoir may flow onto the porous sample collection media.

[0116] In some embodiments, the liquid reservoir (e.g., capsule) may be a separate element from the sample collection device and may be constructed to be received by the housing. The housing may optionally include a receptacle for receiving the liquid capsule. The liquid capsule may be a replacement element with the sample collection device. The liquid reservoir may include a membrane or seal that is removable, breakable, or pierceable. One illustrative metered fluid dose element is commercially available from 3M Company (St. Paul, MN, U.S.A.) under the trade designation CUROS. The housing may include one or more features, such as a piercing element, that the liquid reservoir may be pressed against to rupture, puncture, peel, or otherwise break a seal on the capsule to release the metered dose of liquid. The user may twist and/or press the capsule against the piercing element or other feature to release the metered dose of liquid onto the porous sample collection media.

[0117] Liquid may also be dispensed onto the porous sample collection media from a separate reservoir, such as a dropper.

[0118] The liquid dispensed onto the porous sample collection media may be an aqueous liquid. The liquid may be a buffer solution. The liquid may be an aqueous buffer solution. The liquid may be a saline solution. The liquid may include a surfactant. In some embodiments, the liquid may be an aqueous solution including a surfactant. A “surfactant” is generally understood to mean a molecule that can be added to a solution to reduce the surface tension of the solution. The liquid may be formulated to have a surface tension that facilitates release from the reservoir, as well as flow through or across the porous sample collection media and onto the assay. For example, the surface tension of the liquid may be lower than that of water. The liquid may have a contact angle of greater than 90 degrees when measured on the porous sample collection media. The liquid may be a saline solution including a surfactant. The liquid (for example, a buffer or a saline solution) may include from 0.1 wt-% or more or 0.5 wt-% or more, and up to 1 wt-% or up to 2 wt-% of surfactant.

[0119] The liquid may be provided as a metered dose. The metered dose of liquid may have a volume of 50 microliters or greater, 100 microliters or greater, 150 microliters or greater, 200 microliters or greater, or 250 microliters or greater. The metered dose of liquid may have a volume of 1200 microliters or less, 1000 microliters or less, 750 microliters or less, 500 microliters or less, 400 microliters or less, 300 microliters or less, 250 microliters or less, or 200 microliters or less. The metered dose of liquid may have a volume of 50 microliters to 1000 microliters, 100 microliters to 500 microliters, 100 microliters to 250 microliters. The metered dose may include from 50 microliters to 1000 microliters of buffer.

[0120] In some embodiments, the volume of the metered dose of a liquid is relative to the surface area of the porous sample collection media. The porous sample collection media defines a major surface area and the metered dose of the liquid defines a volume, and the volume divided by the surface area may be in a range from 10 pm/cm² to 400 pm/cm², or from 10 pm/cm² to 250 pm/cm², or from 50 pm/cm² to 150 pm/cm². In some embodiments, the metered dose of liquid defines a volume in a range from 50 microliters to 1000 microliters or 100 microliters to 500 microliters.

[0121] Without wishing to be bound by theory, it is believed that by obtaining a sample that includes dislodged mucosal particulate from the respiratory tract’s epithelial cells, and by using a relatively small volume of liquid to elute the analyte, the sensitivity of the test can be increased as compared to methods that use EBC.

[0122] In some embodiments, the system includes two or more liquid reservoirs, each liquid reservoir being constructed to deliver liquid onto the porous sample collection media. At least one of the two or more liquid reservoirs may contain a liquid different than a liquid contained in another of the two or more liquid reservoirs. The liquid contained in the two or more liquid reservoirs may be different from one another. For example, one liquid reservoir may contain a buffer solution to elute the sample and another liquid reservoir may contain a reagent used to analyze the sample. The liquids from the two or more liquid reservoirs may be released simultaneously or in succession.

[0123] The liquid may be applied onto the loaded porous sample collection media to elute the sample. The liquid may travel through the surface and thickness of the loaded porous sample collection media and flow off of the porous sample collection media carrying with it the target analyte present on the loaded porous sample collection media. This loaded liquid may then be received by the assay and tested for the presence of the target analyte. In some embodiments, the target analyte is virus, bacteria, fungus, pathogen, or other analyte (e.g., protein or cell wall fragment). In some embodiments, the target analyte is LAM.

[0124] In some embodiments, the liquid reservoir 200’ may include an opening and a removable cover 250’ on the opening (see FIG. 5B). In some embodiments, the removable cover 250’ may be positioned between the volume V200’ and the porous sample collection media 120. The removable cover may removably seal the liquid reservoir. The removable cover may be sealed onto the opening of the liquid reservoir by a peelable seal. The peelable seal may be formed, for example, by a heat seal, a suitable adhesive, a gasket, or a combination thereof. In some embodiments, a metered dose of liquid may be released by pulling on a removable cover from an opening on the liquid reservoir. In some embodiments, the removable cover 250’ is a removable tab. In some embodiments, the removable tab forms a pull tab. Exemplary suitable materials for the removable cover or the removable tab, available from the 3M company, include 3M 9792R Single-Coated Aluminum; 3M 9793R SC Polyolefin; 3M 9794R SC Clear PET; 3M 9795R SC Advanced Polyolefin; 3M 9964 SC Clear PET; and 3M 9965 Double-Coated White PET [2 Liners], Suitable adhesives include adhesives that seal the reservoir such that the liquid does not flow out prematurely but allow the cover or the tab to be removed by a user.

[0125] In some embodiments, the liquid may be disposed directly in the liquid reservoir formed by the housing (without incorporating a capsule). The liquid reservoir may be sealed by the removable cover or the removable tab.

Sample Collection System without Assay

[0126] In some embodiments, the sample collection system may not include an assay, or another test method is used in addition to an assay. In such embodiments, the sample collection system may include many of the same features as discussed above. For example, the sample collection system includes a sample receiving element and a sample collection device including a sheet of porous sample collection media and a housing or holder constructed to support the porous sample collection media, as described above.

[0127] Samples from sample collection devices with or without an assay may be transferred and eluted to be analyzed using another assay or test method. This opens up the possibility to use a broader range of analytical methods without limitations of the device, while still being able to benefit from the convenience of the sampling method, the sample type (a respiratory tract sample), and the concentration of the analyte in the porous sample collection media. The analytical methods are not particularly limited, and may be selected based on the target analyte(s), desired detection limit, and the resources available. The target analytes may include virus, bacteria, fungus, pathogen, or other analyte (e.g., protein or cell wall fragment). In embodiments, the target analyte is a virus. The virus may be a coronavirus, rhinovirus, norovirus, influenza virus, adenovirus, adeno-associated virus, varicella-zoster virus, herpesvirus, retrovirus, papillomavirus, enterovirus, arenavirus, or another type of virus. In some embodiments, the virus is present in a respiratory tract sample. In embodiments wherein the target analyte is a coronavirus, it may be SARS-CoV-2 virus (e.g., COVID-19). In some embodiments, the target analyte is bacteria. The bacteria may be, for example, Mycobacterium tuberculosis (Mtb), Streptococcus pneumoniae, Mycoplasma pneumoniae, Haemophilus influenzae, Chlamydophila pneumoniae, Coxiella burnetiid, Moraxella catarrhalis, Histoplasma capsulatum, o Legionella pneumophila. In some embodiments, the target analyte is a fungus. The fungus may be, for example, Cryptococcus neoformans, Aspergillus, Pneumocystis, or endemic fungi. The target analyte may be a disease indicator, such as a protein or cell wall fragment. The test methods that may be used to test the target analyte(s) are not particularly limited and any known methods may be used to test the sample obtained using the sample collection device. Examples of analytical methods that may be used to analyze LAM include immunoassays, separation methods coupled with mass spectrometry (e.g., matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDLTOF) or supercritical fluid chromatography- mass spectrometry (SFC-MS)). Mass-spectrometry may allow the detection of LAM amounts of as low as 25 picograms (pg), while other detection methods may be able to detect in the hundreds of picograms. [0128] In some embodiment, the sample obtained using the sample collection device is analyzed for the presence of LAM (either by using an assay present in the sample collection system or a separate test method) and Mtb (by using any available test methods, such as known laboratory methods).

Additional Features

[0129] The sample collection device or system may include various additional optional features that may make the use of the device or system more convenient, may protect the device or system, may facilitate obtaining a clean sample, may facilitate eluting the sample, may facilitate analyzing the sample, etc.

[0130] In embodiments where the sample collection system includes an assay, the housing may include a test result display window (see, e.g., window 170 in FIGS. 1A, 5A, and window 1170 in FIG. 9A, window 2170 in FIG. 10A, window 3170 in FIG. 11A, and window 4170 in FIG. 12A). The housing may include a viewing window (either transparent material or an opening) in the area of the result display. In some cases, the entire housing may be made of a transparent material. The viewing window 170, 1170, 2170, 3170, 4170 may include one or more markers that help the user to interpret the results of the assay. The one or more markers may include a marker for where the control line will appear on the assay. In an exemplary embodiment, the marker for the control line may include the letter “C.” The one or more markers may include a marker for a positive analysis result of the target analyte(s). In an exemplary embodiment shown in FIGS. 10A and 10B, markers for multiple analytes are included, including “CO” for COVID- 19, “FA” for influenza (“flu”) A, and “FB” for influenza B. In another embodiment, the one or more markers may include a marker for tuberculosis. For example, the one or more markers may include “TB” for tuberculosis in a sample collection system configured for testing for lipoarabinomannan (LAM) to indicate the presence of Mycobacterium tuberculosis (Mtb). In addition, the markers may include a connecting line 2175 (see FIG. 10A) from the top of the housing 2100 to the bottom of the window 2170 to help the user to connect the line on the assay to the correct marker.

[0131] The housing or holder may further include a cover or sealing layer constructed to prevent contamination before or after use of the system. The cover or sealing layer may be removable (for example, may be removed before use). The cover or sealing layer may be closable and/or re- closable (for example, may be closed after use).

[0132] The housing or holder may include a pre-filter or screen disposed in the airflow path in front (upstream) of the porous sample collection media. The screen may be constructed to catch larger particles (larger than the target analyte) and prevent such particles from reaching the porous sample collection media. The exhalation airflow passes through a thickness of the prefilter or screen. The pre-filter or screen at least partially occludes the air flow path. The pre-filter or screen may be a non-woven layer configured to filter out larger particles from the exhalation airflow passing through the pre-filter or screen. In some cases, the pre-filter or screen may be a nonwoven layer that does not have an electrostatic charge. In some embodiments, the pre-filter or screen does not capture significant amounts of the target analyte and instead allows it to be transmitted through the pre-filter or screen. In some embodiments, the pre-filter or screen is made of or includes at least one of a plastic mesh, a woven net, a needle-tacked fibrous web, a knitted mesh, an extruded net, and/or a carded or spunbond coverstock.

[0133] The sample collection device may include a removable and replaceable airflow outlet cover. The airflow outlet cover may be removed from the housing by a user to provide an airflow outlet in the housing. The user may further replace the airflow outlet cover to close the opening and to prevent liquid flow through the opening once the metered dose of liquid is applied onto the porous sample collection media. The airflow outlet cover may be provided as a separate piece detached from the device. The airflow outlet cover may be connected to the device. The airflow outlet cover may be provided as connected to the bottom portion of the housing. For example, the airflow outlet cover may be pivotably, slidably, hingedly, or peelably connected to the bottom portion of the housing.

[0134] The sample collection systems discussed here may include an area for writing or otherwise indicating identifying information, such as a name, initials, account number, or the like.

[0135] The sample collection system of the present disclosure may further include a machine- readable optical label. Such labels may include, for example, a bar code and a QR (quick response) code. The machine-readable optical label may be configured to display the result of the assay. The machine-readable optical label may be used to read and record the result. An electronic reader capable of reading machine-readable optical labels may be used to read and record the result. An electronic reader may be, for example, a smartphone, a tablet, a laptop, or bar code reader, or a QR code reader. The electronic reader may further be used to transmit the result, for example, to a healthcare provider or to a database.

Method of Obtaining Sample, Method of Analyzing Sample

[0136] A method of collecting a sample includes exhaling from one or both nostrils through the porous sample collection media to capture a sample in the porous sample collection media, and to form a loaded porous sample collection media. In embodiments that do not include a sample receiving element as part of the sample collection device, the user may simply hold the porous sample collection media (e.g., by holding the holder) against the nostril and exhale through the porous sample collection media. In embodiments that include a sample receiving element as part of the sample collection device, the method may include exhaling through the sample receiving element (for example, into the first end of the sample receiving element) and through the porous sample collection media. The user may exhale through a single nostril by closing the other nostril, for example, by pressing at the side of the nose, leaving one nostril open. Closing one nostril increases the flow rate through the open nostril. The user may exhale through a single nostril to produce a flow rate of 30 L/min or greater, 40 L/min or greater, 50 L/min or greater, 60 L/min or greater, or 70 L/min or greater. The user may exhale through a single nostril to produce a flow rate of 120 L/min or lower, 110 L/min or lower, 100 L/min or lower, 90 L/min or lower, or 80 L/min or lower.

[0137] The sample may be eluted from the porous sample collection media either in place (while the porous sample collection media remains in the housing), or by first removing the porous sample collection media and placing it into another container, such as a test tube. The sample may be eluted by applying a metered dose of liquid onto the porous sample collection media.

[0138] In embodiments where the sample collection system includes an assay and a liquid reservoir, the metered dose of liquid may be released by at least partially removing (or moving) the removable cover (e.g., removable tab) from the liquid reservoir. Alternatively, the metered dose of liquid may be released by rupturing or piercing the liquid reservoir (e.g., a blister). Releasing the metered dose of liquid onto the porous sample collection media elutes the sample from the loaded porous sample collection media and allows the eluted sample to flow onto the assay. The test result may be observed in the result display of the assay. In some embodiments, the observing of the test result includes observing a positive result if the target analyte is present, or observing a negative result if the target analyte is absent. In some embodiments, the observing of the test result includes observing a positive result if LAM is present, or observing a negative result if LAM is absent. The method may further include reading the result display of the assay using an electronic reader.

[0139] In some embodiments, the housing includes an indicator that indicates whether sufficient airflow has been applied to obtain a sample. The method may include flowing exhalation air from a single nostril into the sample collection device (e.g., through the first open end of the sample receiving element) until the airflow indicator provides a positive indicator of sufficient airflow. The positive indication may be visual, audible, or tactile, or a combination of two or more thereof. The positive indication may include the appearance of an element or color, the position of a marker, or a sound.

[0140] In some embodiments, the housing includes one or more markers that help the user to interpret the results of the assay. The markers may include a connecting line from the top of the housing to the bottom of the test result viewing window to help the user to connect the line on the assay to the correct marker. The method may include observing one or more lines on the assay and comparing the one or more lines to a marker on the housing. For example, the method may include observing a line on the assay and following a connecting line from the line on the assay to a marker on the housing. The marker may include the letter “C” for “control.” The method may include observing another line on the assay and following a connecting line from the line on the assay to a marker on the body, where the marker includes a positive indication of an analyte. The method may include observing another line on the assay and following a connecting line from the line on the assay to a marker on the body, where the marker includes “CO” for COVID-19, “FA” for influenza (“flu”) A, “FB” for influenza B, “TB” for tuberculosis, or another disease indication.

[0141] In embodiments where the sample collection device does not include an assay, the method may include transferring the loaded porous sample collection media with the captured sample to a receptacle. In embodiments where the sample collection device includes a holder, the loaded porous sample collection media with the captured sample may be transferred to a receptacle using the holder. For example, the user may grip the second end of the holder that forms a handle and transfer the loaded porous sample collection media with the captured sample without coming into contact with the porous sample collection media. The method may further include applying a liquid onto the loaded porous sample collection media to form an eluted sample; and testing the eluted sample for presence of the target analyte(s) using a test method. The method may further include applying a liquid onto the loaded porous sample collection media to form an eluted sample; and testing the eluted sample for presence of lipoarabinomannan (LAM) using a test method. The test methods may include a chromatographic method, mass- spectrometry, an immunoassay such as an Enzyme-Linked Immunosorbent Assay (ELISA), a Lateral Flow Assay (LFA), a Vertical Flow Assay (VFA), or any other suitable test method for the targe analyte(s), or a combination of two or more thereof.

[0142] In some embodiments, the nasal exhalation sample is collected along with a urine sample. In some embodiments, the nasal exhalation sample is collected along with a nasal swab sample. In some embodiments, the nasal exhalation sample is collected along with an oral swab sample. The samples may be processed and tested together or separately. The nasal exhalation sample may be collected along with a urine sample and the samples may be processed and tested together or separately. The nasal exhalation sample may be collected along with an oral swab sample and the samples may be processed and tested together or separately. The nasal exhalation sample may be collected along with a nasal swab sample and the samples may be processed and tested together or separately.

[0143] For example, an aliquot (e.g., about 100 pL to about 500 pL, about 100 pL to about 300 pL, or about 100 pL to about 150 pL) of a urine sample may be added into the same container (e g., a vial or test tube) as the nasal exhalation sample. The aliquot of urine sample, alone or mixed with an elution liquid or buffer (as described elsewhere herein), may be added into the same container (e.g., a vial or test tube) as the sample collection media. The aliquot of urine sample, alone or mixed with a buffer (as described elsewhere herein), may be used to elute the nasal exhalation sample from the sample collection media. The aliquot of urine sample may be mixed with an eluted nasal exhalation sample. The urine sample and nasal exhalation sample may be tested together using the same assay or test method. [0144] An eluted swab sample may be mixed with an eluted nasal exhalation sample. The swab sample and nasal exhalation sample may be tested together using the same assay or test method.

Kit

[0145] The sample collection system may be provided as a kit. The kit may include the sample collection system as discussed above, and instructions for collecting a sample and testing the sample using the assay. The instructions may include instructions to exhale from one or both nostrils (e.g., from a single nostril) through the porous sample collection media to capture a sample in the porous sample collection media, and to form a loaded porous sample collection media. In embodiments that do not include a sample receiving element as part of the sample collection device, the instructions may include instructions to hold the porous sample collection media (e.g., by holding the holder) against the nostril and exhale through the porous sample collection media. In embodiments that include a sample receiving element as part of the sample collection device, the instructions may include instructions to exhale through the sample receiving element (for example, into the first end of the sample receiving element) and through the porous sample collection media. The instructions may include instructions to exhale through a single nostril by closing the other nostril, for example, by pressing at the side of the nose, leaving one nostril open. The instructions may include instructions to exhale through a single nostril to produce a flow rate of 30 L/min or greater, 40 L/min or greater, 50 L/min or greater, 60 L/min or greater, or 70 L/min or greater. The instructions may include instructions to exhale through a single nostril to produce a flow rate of 120 L/min or lower, 110 L/min or lower, 100 L/min or lower, 90 L/min or lower, or 80 L/min or lower. The instructions may include instructions to exhale through a single nostril until an airflow indicator indicates a sufficient airflow amount, volume, or pressure. For example, the instructions may include instructions to exhale through a single nostril until an airflow indicator tab (e.g., with a color indicator) appears. The instructions may include instructions to exhale through a single nostril until an airflow indicator marker indicates sufficient airflow. The instructions may include instructions to exhale through a single nostril until an airflow indicator sound indicates sufficient airflow.

[0146] The instructions may further include instructions to move or remove the removable cover or removable tab to release a metered dose of liquid onto the porous sample collection media; and observe a test result in a result display of the assay. The instructions may further include instructions to read a test result display of the assay using an electronic reader.

[0147] The instructions may further include instructions to transfer the loaded porous sample collection media with the captured sample to a receptacle. In embodiments where the sample collection device includes a holder, the loaded porous sample collection media with the captured sample may be transferred to a receptacle using the holder. For example, the instructions may instruct the user to grip the second end of the holder that forms a handle and transfer the loaded porous sample collection media with the captured sample without coming into contact with the porous sample collection media. The instructions may further include instructions to apply a metered dose of liquid onto the loaded porous sample collection media to form an eluted sample; and testing the eluted sample for presence of the target analyte(s) using a test method. The instructions may include instructions to test the eluted sample for presence of lipoarabinomannan (LAM) using a test method.

Embodiments

[0148] The following is a list of exemplary embodiments according to the present disclosure.

[0149] Embodiment 1 is a sample collection system comprising: a sample collection device comprising: a housing; a sample receiving element extending from the housing, the sample receiving element comprising an air inlet constructed to receive an exhalation sample from a single nostril; a porous sample collection media disposed within the housing; an airflow path extending from the air inlet through the porous sample collection media; and an assay constructed to receive an eluted sample from the porous sample collection media and to detect presence of a target analyte in the eluted sample.

[0150] Embodiment 2 is the sample collection system of embodiment 1, comprising: a liquid reservoir in fluid communication with the porous sample collection media, constructed to direct liquid onto the porous sample collection media, the liquid reservoir comprising: a volume housing the liquid; an opening; and a cover on the opening configured to be removed, ruptured, or pierced, optionally wherein the liquid reservoir is immediately adjacent to the air inlet.

[0151] Embodiment 3 is the sample collection system of embodiment 2, wherein the cover is positioned between the volume and the porous sample collection media. [0152] Embodiment 4 is the sample collection system of embodiment 2 or 3, wherein the cover is a removable tab.

[0153] Embodiment 5 is the sample collection system of embodiment 3, wherein the removable tab is a pull tab.

[0154] Embodiment 6 is the sample collection system of any one of embodiments 2 to 5, wherein the porous sample collection media defines a surface area, and the volume of the liquid reservoir divided by the surface area is in a range from 10 pm/cm² to 400 pm/cm², or from 10 pm/cm² to 250 pm/cm², or from 50 pm/cm² to 150 pm/cm².

[0155] Embodiment 7 is the sample collection system of any one of embodiments 2 to 6, wherein the volume is in a range of 50 microliters to 1000 microliters.

[0156] Embodiment 8 is the sample collection system of any one of embodiments 2 to 7, wherein the liquid reservoir houses from 50 microliters to 1000 microliters of the liquid.

[0157] Embodiment 9 is the sample collection system of any one of embodiments 2 to 8, wherein the metered dose of liquid has a volume of 50 microliters or greater, 100 microliters or greater, 150 microliters or greater, 200 microliters or greater, or 250 microliters or greater.

[0158] Embodiment 10 is the sample collection system of any one of embodiments 2 to 9, wherein the metered dose of liquid has a volume of 1200 microliters or less, 1000 microliters or less, 750 microliters or less, 500 microliters or less, or 400 microliters or less.

[0159] Embodiment 11 is the sample collection system of any one of embodiments 2 to 10, wherein the metered dose of liquid has a volume of 50 microliters to 1000 microliters or 100 microliters to 500 microliters.

[0160] Embodiment 12 is the sample collection system of any one of the preceding embodiments, wherein the housing comprises a first part coupled with a second part.

[0161] Embodiment 13 is the sample collection system of embodiment 12, wherein the air inlet is formed in the first part.

[0162] Embodiment 14 is the sample collection system of embodiment 12 or 13, wherein an air outlet is formed in the first part. [0163] Embodiment 15 is the sample collection system of any one of embodiments 12 to 14, wherein the first part is a top part and the second part is a bottom part.

[0164] Embodiment 16 is the sample collection system of any one of embodiments 12 to 14, wherein the assay is disposed between the first and second parts.

[0165] Embodiment 17 is the sample collection system of any one of embodiments 12 to 14, wherein the first part includes a result viewing window through which an assay result area is viewable.

[0166] Embodiment 18 is the sample collection system of any one of embodiments 12 to 14, wherein the first and second parts comprise corresponding coupling features.

[0167] Embodiment 19 is the sample collection system of embodiment 18, wherein the first part and second part are connected by a snap fit closure, and optionally wherein the snap fit closure is tamper evident.

[0168] Embodiment 20A is the sample collection system of any one of the preceding embodiments, wherein the housing comprises an airflow indicator constructed to provide an indication of a predetermined amount of airflow.

[0169] Embodiment 20B is the sample collection system of embodiment 20A, wherein the indication is visual, audible, or tactile, or a combination thereof.

[0170] Embodiment 20C is the sample collection system of embodiment 20B, wherein the airflow indicator comprises a spring mechanism constructed to provide resistance to airflow applied to the air inlet.

[0171] Embodiment 20D is the sample collection system of embodiment 20C, wherein the airflow indicator comprises a tab constructed to extend through an aperture in the housing upon application of airflow exceeding the resistance of the spring mechanism.

[0172] Embodiment 20E is the sample collection system of embodiment 20A, wherein the airflow indicator comprises a marker constructed to slide along an indicator track comprising a first portion that indicates insufficient airflow and a second portion that indicates sufficient airflow, and wherein the marker is constructed to move from a first position within the first portion to a second position within the second portion when sufficient amount of airflow has been applied to the air inlet. [0173] Embodiment 20F is the sample collection system of embodiment 20 A, wherein the airflow indicator comprises a whistle, optionally wherein the whistle is disposed within the sample receiving element.

[0174] Embodiment 21 A is the sample collection system of any one of the preceding embodiments, wherein the porous sample collection media comprises a nonwoven filtration layer having an electrostatic charge.

[0175] Embodiment 21B is the sample collection system of embodiment 21A, wherein the nonwoven filtration layer is hydrophobic.

[0176] Embodiment 22 is the sample collection system of any one of embodiments 2 to 21B, wherein the liquid is an aqueous solution comprising a surfactant.

[0177] Embodiment 23 is the sample collection system of any one of the preceding embodiments, comprising two or more liquid reservoirs, each liquid reservoir constructed to deliver liquid onto the porous sample collection media, optionally wherein at least one of the two or more liquid reservoirs contains a liquid being different than a liquid contained in another of the two or more liquid reservoirs.

[0178] Embodiment 24 is the sample collection system of any one of the preceding embodiments, further comprising a pre-fdter fixed within the housing and along the airflow path and between the air inlet and the porous sample collection media.

[0179] Embodiment 25 A is the sample collection system of any one of the preceding embodiments, wherein the assay is constructed to detect presence of lipoarabinomannan (LAM) in a collected sample.

[0180] Embodiment 25B is the sample collection system of embodiment 25 A, wherein the housing comprises a test result display window and a marker adjacent the test result display window marking a position of a positive test result for tuberculosis when the presence of lipoarabinomannan (LAM) is detected.

[0181] Embodiment 25C is the sample collection system of embodiment 25B, wherein the housing comprises a connecting line from the marker to the position of test result on the assay. [0182] Embodiment 26 is the sample collection system of any one of the preceding embodiments, wherein the assay is a lateral flow assay, a vertical flow assay, or a colorimetric indicator.

[0183] Embodiment 27A is the sample collection system of any one of the preceding embodiments, wherein the housing comprises a test result display window.

[0184] Embodiment 27B is the sample collection system of embodiment 27A, wherein the housing comprises a marker adjacent the test result display window marking a position of a positive test result on the assay.

[0185] Embodiment 28 is the sample collection system of any one of the preceding embodiments, wherein the assay has a LAM detection limit of 10 picograms/milliliters or lower.

[0186] Embodiment 29 is a sample collection system comprising: a sample receiving element with a tubular body comprising: a first open end constructed to receive an exhalation sample from a single nostril; a second open end; and an air channel extending between the first open end and the second open end; and a sample collection device comprising: a sheet of porous sample collection media; a holder comprising a first end coupled with and constructed to support the porous sample collection media, and a second end forming a handle, wherein the sheet of porous sample collection media is removably received in the sample receiving element and arranged to receive a through-flow of the exhalation sample.

[0187] Embodiment 30 is the sample collection system of any one of the preceding embodiments, wherein the tubular body of the sample receiving element comprises a frustoconical shape extending from the first open end toward the second open end, wherein the frustoconical shape has its smallest diameter at the first open end.

[0188] Embodiment 31 is the sample collection system of embodiment 30, wherein the second open end of the tubular body of the sample receiving element is constructed for receiving a breath sample from a user’s mouth.

[0189] Embodiment 32 is a method of collecting and testing a sample, the method comprising: flowing exhalation air from a single nostril through a porous sample collection media to form a captured sample, the porous sample collection media being disposed within a sample collection device, the sample collection device comprising a sample receiving element with a tubular body comprising: a first open end constructed to receive the exhalation air from a single nostril; a second open end; and an air channel extending between the first open end and the second open end; releasing a metered dose of liquid onto the porous sample collection media from a liquid reservoir, the metered dose of liquid eluting the captured sample onto an assay disposed within the sample collection device housing, the assay being constructed to receive an eluted sample from the porous sample collection media and to detect presence of a target analyte in the eluted sample; and observing a test result on the assay.

[0190] Embodiment 33 is the method of embodiment 32, wherein the liquid reservoir is disposed within the sample collection device housing.

[0191] Embodiment 34 is the method of embodiment 32 or 33, wherein the metered dose of liquid is released by pulling on a removable cover from an opening on the liquid reservoir.

[0192] Embodiment 35 is the method of any one of embodiments 32 to 34, wherein the removable cover is a removable tab.

[0193] Embodiment 36A is the method of embodiment 35, wherein the removable tab is a pull tab.

[0194] Embodiment 36B is the method of embodiment 36A, wherein the pulling on the pull tab unseals an opening on the liquid reservoir.

[0195] Embodiment 37 is the method of embodiment 32 or 33, wherein the metered dose of liquid is released by piercing a cover on the liquid reservoir.

[0196] Embodiment 38 is the method of any one of embodiments 32 to 37, wherein the metered dose comprises from 50 microliters to 1000 microliters of buffer. The metered dose of liquid may have a volume of 50 microliters or greater, 100 microliters or greater, 150 microliters or greater, 200 microliters or greater, or 250 microliters or greater. The metered dose of liquid may have a volume of 1200 microliters or less, 1000 microliters or less, 750 microliters or less, 500 microliters or less, or 400 microliters or less. The metered dose of liquid may have a volume of 50 microliters to 1000 microliters or 100 microliters to 500 microliters. The metered dose may include from 50 microliters to 1000 microliters of buffer. [0197] Embodiment 39 is the method of any one of embodiments 32 to 38, wherein flowing exhalation air comprises blowing into a mouthpiece or nosepiece on the sample collection device.

[0198] Embodiment 40A is the method of any one of embodiments 32 to 39, wherein the observing of the test result comprises observing a positive result if LAM is present, or observing a negative result if LAM is absent.

[0199] Embodiment 40B is the method of embodiment 40 A, wherein the observing of the test result further comprises observing a marker for tuberculosis adjacent the positive test result for LAM.

[0200] Embodiment 41 is the method of any one of embodiments 32 to 40, wherein the assay has a LAM detection limit of 10 picograms/milliliter or lower.

[0201] Embodiment 42 is a method of collecting and testing a sample, the method comprising: flowing exhalation air from a single nostril through a porous sample collection media to form a captured sample, the porous sample collection media being disposed within a sample collection device comprising a sample receiving element with a tubular body comprising: a first open end constructed to receive the exhalation air from the single nostril; a second open end; and an air channel extending between the first open end and the second open end; transferring the porous sample collection media and the captured sample to a receptacle using a holder comprising a first end coupled with and constructed to support the porous sample collection media, and a second end forming a handle; applying a liquid onto the porous sample collection media and the captured sample to form an eluted sample; and testing the eluted sample for presence of target analyte using a test method.

[0202] Embodiment 43 is the method of embodiment 42, wherein the test method comprises a chromatographic method.

[0203] Embodiment 44 is the method of embodiment 42, wherein the test method comprises an Enzyme-Linked Immunosorbent Assay (ELISA).

[0204] Embodiment 45 is the method of embodiment 42, wherein the test method comprises a Lateral Flow Assay or a Vertical Flow Assay. [0205] Embodiment 46 is the method of any one of embodiments 32 to 45, further comprising reading a QR code on the sample collection device.

[0206] Embodiment 47 is the method of any one of embodiments 32 to 46 further comprising collecting a second sample by collecting urine and testing the urine for presence of the target analyte.

[0207] Embodiment 48 is the method of embodiment 47, wherein the second sample is combined with the nasal exhalation sample.

[0208] Embodiment 49 is the method of embodiment 47, wherein the second sample is used to elute the nasal exhalation sample from the sample collection media.

[0209] Embodiment 50 is the sample collection system or method of any one of embodiments 1 to 49, wherein the target analyte comprises a disease indicator or pathogen or both.

[0210] Embodiment 51 is the sample collection system or method of any one of embodiments 1 to 50, wherein the target analyte comprises virus, bacteria, fungus, pathogen, protein, cell wall fragment, lipoarabinomannan (LAM), or a combination of two or more thereof.

[0211] Embodiment 52 is the sample collection system or method of any one of embodiments 1 to 51, wherein the target analyte comprises LAM.

[0212] Embodiment 53 is the sample collection system or method of any one of embodiments 1 to 52, wherein the target analyte comprises LAM and Mtb.

[0213] Embodiment 54 is a kit comprising the sample collection system of any one of embodiments 1 to 31 and instructions for collecting a sample and testing the sample using an assay.

[0214] Embodiment 55 is the kit of embodiment 54, wherein the instructions include instructions to: exhale from a nostril (e.g., from a single nostril) through the porous sample collection media to capture a sample in the porous sample collection media, and to form a loaded porous sample collection media.

[0215] Embodiment 56 is the kit of embodiment 54 or 55, wherein the instructions include instructions to: release a metered dose of liquid onto the porous sample collection media; and observe a test result in a result display of the assay. [0216] Embodiment 57 is the kit of any one of embodiments 54 to 56, wherein the instructions include instructions to transfer the loaded porous sample collection media with the captured sample to a receptacle, optionally using the holder, to apply a metered dose of liquid onto the loaded porous sample collection media to form an eluted sample; and to test the eluted sample for presence of the target analyte(s) using a test method.

[0217] Embodiment 58 is the kit of any one of embodiments 54 to 57, wherein the instructions include instructions to observe an airflow indicator to indicate sufficient airflow.

[0218] All references and publications cited herein are expressly incorporated herein by reference in their entirety into this disclosure, except to the extent they may directly contradict this disclosure. Although specific embodiments have been illustrated and described herein, it will be appreciated by those of ordinary skill in the art that a variety of alternate and/or equivalent implementations can be substituted for the specific embodiments shown and described without departing from the scope of the present disclosure. It should be understood that this disclosure is not intended to be unduly limited by the illustrative embodiments and examples set forth herein and that such examples and embodiments are presented by way of example only with the scope of the disclosure intended to be limited only by the claims set forth here.

Claims

1. A sample collection system comprising: a sample collection device comprising: a housing; a sample receiving element extending from the housing, the sample receiving element comprising an air inlet constructed to receive an exhalation sample from a single nostril; a porous sample collection media disposed within the housing; and an airflow path extending from the air inlet through the porous sample collection media; and an assay constructed to receive an eluted sample from the porous sample collection media and to detect presence of a target analyte in the eluted sample.

2. The sample collection system of claim 1, comprising: a liquid reservoir in fluid communication with the porous sample collection media, constructed to direct liquid onto the porous sample collection media, the liquid reservoir comprising: a volume housing the liquid; an opening; and a cover on the opening configured to be removed, ruptured, or pierced, optionally wherein the liquid reservoir is immediately adjacent to the air inlet.

3. The sample collection system of claim 2, wherein the removable cover is positioned between the volume and the porous sample collection media.

4. The sample collection system of claim 2 or 3, wherein the removable cover comprises a pull tab.

5. The sample collection system of any one of claims 2 to 4, wherein the porous sample collection media defines a surface area, and the volume of the liquid reservoir divided by the surface area is in a range from 10 pm/cm² to 400 pm/cm², or from 10 pm/cm² to 250 pm/cm².

6. The sample collection system of any one of claims 2 to 5, wherein the volume is in a range of 50 microliters to 1000 microliters.

7. The sample collection system of any one of the preceding claims, wherein the porous sample collection media comprises a nonwoven filtration layer having an electrostatic charge.

8. The sample collection system of claim 7, wherein the nonwoven filtration layer is hydrophobic.

9. The sample collection system of claim 2 or 8, wherein the liquid is an aqueous solution comprising a surfactant.

10. The sample collection system of any one of the preceding claims, wherein the assay is constructed to detect presence of lipoarabinomannan (LAM) in a collected sample.

11. The sample collection system of any one of the preceding claims, wherein the assay is a lateral flow assay, a vertical flow assay, or a colorimetric indicator.

12. The sample collection system of any one of the preceding claims, wherein the assay has a LAM detection limit of 10 picograms/milliliters or lower.

13. The sample collection system of any one of the preceding claims, wherein the housing comprises an airflow indicator constructed to provide an indication of a predetermined amount of airflow.

14. A sample collection system comprising: a sample receiving element with a tubular body, the sample receiving element comprising: a first open end constructed to receive an exhalation sample from a single nostril; a second open end; and an air channel extending between the first open end and the second open end; and a sample collection device comprising: a sheet of porous sample collection media; and a holder comprising a first end coupled with and constructed to support the porous sample collection media, and a second end forming a handle, wherein the sheet of porous sample collection media is removably received in the sample receiving element and arranged to receive a through-flow of the exhalation sample.

15. The sample collection system of claim 14, wherein the tubular body of the sample receiving element comprises a wall having a frustoconical shape having its smallest diameter adjacent the air inlet.

16. The sample collection system of claim 14, wherein the tubular body of the sample receiving element comprises a slot constructed to receive the porous sample collection media.

17. The sample collection system of claim 14, wherein the second open end of the tubular body of the sample receiving element is constructed for receiving a breath sample from a user’s mouth.

18. A method of collecting and testing a sample, the method comprising: flowing exhalation air from a single nostril through a porous sample collection media to form a captured sample, the porous sample collection media being disposed within a sample collection device, the sample collection device comprising a housing and a sample receiving element with a tubular body, the sample receiving element comprising: a first open end constructed to receive the exhalation air from a single nostril; a second open end; and an air channel extending between the first open end and the second open end; releasing a metered dose of liquid onto the porous sample collection media from a liquid reservoir, the metered dose of liquid eluting the captured sample onto an assay disposed within the sample collection device housing, the assay being constructed to receive an eluted sample from the porous sample collection media and to detect presence of a target analyte in the eluted sample; and observing a test result on the assay.

19. The method of claim 18, wherein the liquid reservoir is disposed within the sample collection device housing.

20. The method of claim 18 or 19, wherein the metered dose of liquid is released by removing a removable tab from an opening on the liquid reservoir.

21 . The method of any one of claims 18 to 20, wherein the metered dose comprises from 50 microliters to 1000 microliters of buffer.

22. The method of any one of claims 19 to 21, wherein the observing of the test result comprises observing a positive result if lipoarabinomannan (LAM) is present, or observing a negative result if LAM is absent.

23. The method of any one of claims 19 to 22, wherein the assay has a LAM detection limit of 10 picograms/milliliter or lower.

24. The method of any one of claims 19 to 23, wherein the housing comprises an airflow indicator, and wherein the method comprises flowing exhalation air from a single nostril through the first open end of the sample receiving element until the airflow indicator provides a positive indicator of sufficient airflow.

25. The method of any one of claims 19 to 24, wherein the observing of the test result comprises observing one or more lines on the assay and comparing the one or more lines to a marker on the housing.

26. A method of collecting and testing a sample, the method comprising: flowing exhalation air from a single nostril through a porous sample collection media to form a captured sample, the porous sample collection media being disposed within a sample collection device comprising a sample receiving element with a tubular body, the sample receiving element comprising: a first open end constructed to receive the exhalation air from the single nostril; a second open end; and an air channel extending between the first open end and the second open end; transferring the porous sample collection media and the captured sample to a receptacle using a holder comprising a first end coupled with and constructed to support the porous sample collection media, and a second end forming a handle; applying a liquid onto the porous sample collection media and the captured sample to form an eluted sample; and testing the eluted sample for presence of a target analyte using a test method.

27. The method of claim 26, wherein the test method comprises a chromatographic method, a mass spectrometry method, an immunoassay, or a combination thereof.

28. The method of claim 27, wherein the test method comprises an Enzyme-Linked Immunosorbent Assay (ELISA).

29. The method of claim 27, wherein the test method comprises a Lateral Flow Assay or a Vertical Flow Assay.

30. The sample collection system or method of any one of claims 1 to 29, wherein the target analyte comprises a disease indicator or pathogen or both.

31. The sample collection system or method of any one of claims 1 to 30, wherein the target analyte comprises virus, bacteria, fungus, pathogen, protein, cell wall fragment, lipoarabinomannan (LAM), or a combination of two or more thereof.

32. The sample collection system or method of any one of claims 1 to 31, wherein the target analyte comprises LAM.

33. The sample collection system or method of any one of claims 1 to 32, wherein the target analyte comprises LAM and Mtb.