WO2021243035A1

WO2021243035A1 - Non-invasive exhaled breath (eb) collection apparatus

Info

Publication number: WO2021243035A1
Application number: PCT/US2021/034543
Authority: WO
Inventors: Hossein KAVEHPOUR; Ali Alshehri; Jeffrey Ruberti; Jonathan ROTHSTEIN; Robert N. CANDLER; Nasim Annabi
Original assignee: The Regents Of The University Of California; Northeastern University; University Of Massachusetts Amherst
Priority date: 2020-05-27
Filing date: 2021-05-27
Publication date: 2021-12-02

Abstract

Provided herein are apparatuses, kits, and methods, for detecting pathogens contained within human breath using a single-use, disposable sample collection device. The apparatus includes a housing having a base and a cover defining a sample collection chamber. A tube extends through the port such that the distal opening is in fluid communication with the sample collection chamber. The proximal opening is configured to receive a gaseous sample containing moisture and direct the gaseous sample to a sample collection surface on the base. The apparatus further includes a cooling device configured to cool the sample collection surface and thereby condense at least a portion of the moisture on the sample collection surface. The apparatus further includes a sample collection material that is a hydrogel disposed on the sample collection surface. The hydrogel is a modified gelatin functionalize with an enzyme (e.g., ACE2).

Description

NON-INVASIVE EXHALED BREATH (EB) COLLECTION APPARATUS

CROSS-REFERENCE TO RELATED APPLICATION [0001] This application claims the benefit of U.S. Provisional Patent Application No. 63/030,767, filed on May 27, 2020, which is hereby incorporated by reference in its entirety.

NOTICE OF MATERIAL SUBJECT TO COPYRIGHT PROTECTION [0002] A portion of the material in this patent document is subject to copyright protection under the copyright laws of the United States and of other countries. The owner of the copyright rights has no objection to the facsimile reproduction by anyone of the patent document or the patent disclosure, as it appears in the United States Patent and Trademark Office publicly available file or records, but otherwise reserves all copyright rights whatsoever. The copyright owner does not hereby waive any of its rights to have this patent document maintained in secrecy, including without limitation its rights pursuant to 37 C.F.R. § 1.14.

BACKGROUND

[0003] The rapid spread and virulence of COVID-19 has exposed a critical need for both individual patient point of care and environmental sampling and testing. To protect society from the current wave of illnesses and the potential second (possibly deadlier wave), efficient sampling and rapid testing should be deployed into critical infrastructure including food processing plants, hospitals, dental offices, essential government offices, airports, etc. The following is a list of qualities of a testing system for a viral pathogen: (a) the test system should provide fast, efficient patient testing taking samples directly from the lungs where the virus is most abundant; (b) the test system should provide a non-invasive test since the high-volume testing will involve children, senior citizens, mental health patients, and many others who cannot tolerate swab tests; (c) swab tests are known to cause secondary infection for high percentage of the sampling cases (d) the gold standard outcome of testing would be monitoring for both virus and antibodies on the same sampling; (e) health care providers need a way to monitor viral load in lungs directly to better understand when patients are no longer contagious to quickly gauge the efficacy of different treatments; (f) a system is needed for environmental monitoring to determine contamination in rooms and surfaces by continuously sampling the air and the aerosols that settle onto counters, floors and other horizontal surfaces; (g) a system is needed for providing data to understand amount of virus in exhaled breath to better understand necessary distancing and danger of infection in the absence of coughs and sneezes.

[0004] Current patient sampling is performed by deep nasal swabbing and there is no known system capable of sampling the environment for COVID-19. It has been shown that viruses can be retrieved from the exhaled breath, by analyzing HEPA filters described in "Influenza virus in human exhaled breath: an observational study" by Fabian, et al. Additionally, it has been shown that viruses can be retrieved from the exhaled breath, by analyzing an Exhaled Breath Condensation (EBC) technique described in "Infectious virus in exhaled breath of symptomatic seasonal influenza cases from a college community" by Yan, et al. Existing EBC may take 30 minutes to an hour of breathing through a tube in order to collect sufficient samples as described in "Exhaled breath condensate: methodological recommendations and unresolved questions." by Horvath et al.

BRIEF SUMMARY

[0005] The purpose and advantages of the disclosed subject matter will be set forth in and apparent from the description that follows, as well as will be learned by practice of the disclosed subject matter. Additional advantages of the disclosed subject matter will be realized and attained by the methods and systems particularly pointed out in the written description and claims hereof, as well as from the appended drawings.

[0006] To achieve these and other advantages and in accordance with the purpose of the disclosed subject matter, as embodied and broadly described, the disclosed subject matter includes an apparatus including a housing having a base and a cover. The base has a sample collection surface and the cover has a port and substantially encloses the sample collection surface thereby defining a sample collection chamber. The apparatus further includes a tube extending from a proximal end to a distal end, where the proximal end has a proximal opening and the distal end has a distal opening. The tube extends through the port such that the distal opening is in fluid communication with the sample collection chamber. The proximal opening is configured to receive a gaseous sample containing moisture and direct the gaseous sample to the sample collection surface. The apparatus further includes a cooling device configured to contact the base and cool the sample collection surface to thereby condense at least a portion of the moisture on the sample collection surface. The apparatus further includes a sample collection material disposed on the sample collection surface. The sample collection material includes a hydrogel configured to absorb the condensed moisture.

[0007] In various embodiments, an exhaled breath sample collection device is provided including a base having a sample collection surface. The sample collection surface is configured to receive a gaseous sample containing moisture and cool the gaseous sample to thereby condense at least a portion of the moisture on the sample collection surface. The device further includes a sample collection material disposed on the sample collection surface. The sample collection material includes a hydrogel configured to absorb the condensed moisture. The device may further include a cover having a port and the cover may substantially enclose the sample collection surface thereby defining a sample collection chamber. The device may further includes a tube extending from a proximal end to a distal end, where the proximal end has a proximal opening and the distal end has a distal opening. The tube may extend through the port such that the distal opening is in fluid communication with the sample collection chamber.

[0008] In various embodiments, a method for forming an exhaled breath sample collection device is provided where a mask is applied to a first portion of a substrate such that a second portion of the substrate does not include the mask. A hydrogel solution is applied to the second portion. The hydrogel solution includes a hydrogel and a photoinitiator. The hydrogel solution is photocrosslinked. The mask is then removed.

[0009] In various embodiments, a method for detecting a pathogen contained within breath of a human is provided where a gaseous sample of human breath containing moisture is received at a sample collection surface on a base. The sample collection surface is cooled to a temperature lower than the dew point of the moisture in the gaseous sample thereby condensing the moisture on the sample collection surface. At least a portion of the condensed moisture is collected at a sample collection material disposed on the sample collection surface. The sample collection material includes a hydrogel.

[0010] In various embodiments, an apparatus is provided including a housing having a base and a cover. The base has a sample collection surface and the cover has a port and substantially encloses the sample collection surface thereby defining a sample collection chamber. The apparatus further includes a tube extending from a proximal end to a distal end, where the proximal end has a proximal opening and the distal end has a distal opening. The tube extends through the port such that the distal opening is in fluid communication with the sample collection chamber. The proximal opening is configured to receive a gaseous sample containing moisture and direct the gaseous sample to the sample collection surface. The apparatus further includes a cooling device configured to contact the base and cool the sample collection surface to thereby condense at least a portion of the moisture on the sample collection surface. The apparatus further includes one or more microfluidic channels disposed within the base, the one or more microfluidic channels being configured to collect the condensed moisture.

[0011] In various embodiments, a method for detecting a virus contained within breath of a human is provided where a gaseous sample of human breath is received containing moisture at a sample collection surface. The sample collection surface is cooled to a temperature lower than the dew point of the moisture in the gaseous sample thereby condensing the moisture on the sample collection surface. At least a portion of the condensed moisture is collected within a microfluidic channel formed in the sample collection surface.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS [0012] The objects, features and advantages of the devices, systems, and methods described herein will be apparent from the following description of particular embodiments thereof, as illustrated in the accompanying drawings. The drawings are not necessarily to scale, emphasis instead being placed upon illustrating the principles of the devices, systems, and methods described herein.

[0013] Fig. 1A illustrates a diagram showing a general method for collecting samples of condensed air moisture for polymerase chain reaction (PCR) analysis of virus presence in accordance with an embodiment of the present disclosure. Fig. IB illustrates a diagram showing a general method for continuous dropwise concentration (CDC) and retained exhaled breath condensate (EBC) for polymerase chain reaction (PCR) analysis of virus presence in accordance with an embodiment of the present disclosure.

[0014] Fig. 2A illustrates a schematic diagram illustrating sample collection in accordance with an embodiment of the present disclosure. Fig. 2B illustrates a sample collection surface having condensed moisture thereon in accordance with an embodiment of the present disclosure.

[0015] Fig. 3A illustrates an exhaled droplet condensation collection apparatus in accordance with an embodiment of the present disclosure. Fig. 3B illustrates an exhaled droplet condensation collection apparatus in accordance with an embodiment of the present disclosure.

[0016] Figs. 4A-4B illustrate a comparison of an exhaled breath collection system (Fig. 4A) and an embodiment of an apparatus according to the present disclosure (Fig. 4B) in accordance with an embodiment of the present disclosure.

[0017] Fig. 5 illustrates a system diagram of baseline use of an exhaled breath collection system in accordance with an embodiment of the present disclosure.

[0018] Fig. 6 illustrates a system diagram illustrating a point-of-care system with fluorescence in situ hybridization (FISH) developer and scanning in accordance with an embodiment of the present disclosure.

[0019] Fig. 7 illustrates a flow diagram of a system workflow for sample collection and analysis in accordance with an embodiment of the present disclosure.

[0020] Fig. 8A illustrates a chemical reaction for conjugation of ACE2 to hydrogel 1HNMR spectra of hydrogel (10% and 20% (w/v)) formed at varying visible light exposure times in accordance with an embodiment of the present disclosure. Fig. 8B illustrates a coating of device with ACE2-hydrogel in accordance with an embodiment of the present disclosure.

[0021] Figs. 9A-9E illustrate a method for performing FISH using condensed breath in accordance with an embodiment of the present disclosure.

[0022] Figs. 10A-10E illustrate a method for performing rtPCR using condensed breath in an apparatus using a swab for moisture collection in accordance with an embodiment of the present disclosure.

[0023] Figs. 11A-11F illustrate a method for performing rtPCR using condensed breath in an apparatus having microfluidic channels in accordance with an embodiment of the present disclosure.

[0024] Figs. 12A-12D illustrate a method for performing FISH using condensed breath in an apparatus having microfluidic channels in accordance with an embodiment of the present disclosure.

[0025] Fig. 13 illustrates an experimental setup used for condensate extraction from a humid air jet in accordance with an embodiment of the present disclosure.

[0026] Fig. 14 illustrates regular camera photographs of the breath figures at varying heights-to-diameter ratios and jet Reynolds numbers in accordance with an embodiment of the present disclosure. [0027] Fig. 15 illustrates an infrared camera pictures of droplet formation at different surface temperature and different times in accordance with an embodiment of the present disclosure.

[0028] Fig. 16 illustrates a graph showing examples of human expiratory flow rate in accordance with an embodiment of the present disclosure.

[0029] Fig. 17 illustrates a diagram of humid-air jet impingement in accordance with an embodiment of the present disclosure.

[0030] Figs. 18A-18B illustrate graphical correlation of the power law constants K and a at various conditions for local Nusselt number in accordance with an embodiment of the present disclosure.

[0031] Fig. 19 illustrates a control volume (CV) on which thermodynamic analysis may be performed in accordance with an embodiment of the present disclosure.

[0032] Fig. 20 illustrates an exemplary system for detecting viral presence in accordance with an embodiment of the present disclosure.

DETAILED DESCRIPTION

[0033] As used herein, the singular terms "a," "an," and "the" may include plural referents unless the context clearly dictates otherwise. Reference to an object in the singular is not intended to mean "one and only one" unless explicitly so stated, but rather "one or more." [0034] As used herein, the term "set" refers to a collection of one or more objects. Thus, for example, a set of objects can include a single object or multiple objects.

[0035] As used herein, the terms "substantially" and "about" are used to describe and account for small variations. When used in conjunction with an event or circumstance, the terms can refer to instances in which the event or circumstance occurs precisely as well as instances in which the event or circumstance occurs to a close approximation. When used in conjunction with a numerical value, the terms can refer to a range of variation of less than or equal to ± 10% of that numerical value, such as less than or equal to ±5%, less than or equal to ±4%, less than or equal to ±3%, less than or equal to ±2%, less than or equal to ±1 %, less than or equal to ±0.5%, less than or equal to ±0.1 %, or less than or equal to ±0.05%. For example, "substantially" aligned can refer to a range of angular variation of less than or equal to ±10°, such as less than or equal to ±5°, less than or equal to ±4°, less than or equal to ±3°, less than or equal to ±2°, less than or equal to ±1°, less than or equal to ±0.5°, less than or equal to ±0.1°, or less than or equal to ±0.05°. [0036] Additionally, amounts, ratios, and other numerical values may sometimes be presented herein in a range format. It is to be understood that such range format is used for convenience and brevity and should be understood flexibly to include numerical values explicitly specified as limits of a range, but also to include all individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly specified. For example, a ratio in the range of about 1 to about 200 should be understood to include the explicitly recited limits of about 1 and about 200, but also to include individual ratios such as about 2, about 3, and about 4, and sub-ranges such as about 10 to about 50, about 20 to about 100, and so forth.

[0037] Although the description herein contains many details, these should not be construed as limiting the scope of the disclosure but as merely providing illustrations of some of the presently preferred embodiments. Therefore, it will be appreciated that the scope of the disclosure fully encompasses other embodiments which may become obvious to those skilled in the art.

[0038] All structural and functional equivalents to the elements of the disclosed embodiments that are known to those of ordinary skill in the art are expressly incorporated herein by reference and are intended to be encompassed by the present claims.

Furthermore, no element, component, or method step in the present disclosure is intended to be dedicated to the public regardless of whether the element, component, or method step is explicitly recited in the claims. No claim element herein is to be construed as a "means plus function" element unless the element is expressly recited using the phrase "means for". No claim element herein is to be construed as a "step plus function" element unless the element is expressly recited using the phrase "step for".

[0039] This disclosure describes apparatuses, systems, kits, and methods for collecting samples of condensed air moisture for viral load level diagnosis. In various embodiments, samples can be collected from a patient in less than a minute. For example, humid air could be sampled from a patient’s breath or from environmental ( e.g ., room) air. In various embodiments, the condensed moisture may be transported to, for example, a polymerase chain reaction (PCR) system for analysis of virus presence.

[0040] This disclosure also describes a condensation mechanism that has experimentally been found to be significantly more efficient than state-of-the art dehumidification systems. In one embodiment, the basic principle of the mechanism is to direct the patient's breath or humid air such that it impinges on a surface that is cooled down to a desired temperature lower than the dew point of the incoming air. For example, a patient could blow into a tube that directs the patient's breath toward the surface as one or more jet of air. In various embodiments, because the surface is cooled below the dew point of the incoming air, a breath figure (BF) spot appears very quickly ( e.g ., almost instantaneously). In various embodiments, this BF spot includes micron-sized water droplets. In various embodiments, as the user breathes through the tube and the droplets grow, the shearing effects of the jet act to push them radially outward to an equilibrium radial location. In various embodiments, the water droplets (that may contain a target, e.g., virus, bacteria, fungus, etc.) may be collected by, for example, either flushing the surface with a sterile liquid (e.g, sterile water) or using a wicking material to absorb the condensed droplets. In various embodiments, the liquid droplets can then be transported for analysis (e.g, PCR, immunofluorescence, staining, etc.).

[0041] This disclosure also describes an apparatus for carrying out the foregoing procedures. In various embodiments, the apparatus may include (a) a collection surface that optionally may be treated with a hydrophobic material, (b) a tube, which is preferably disposable, that directs a patient's exhaled breath to impinge on the collection surface, (c) means for cooling the collection surface (e.g., an thermoelectric or Peltier plate thermally coupled or pasted to the underside of the collection surface) to provide the cooling capacity required for condensation, (d) a cover or enclosure coupled to the tube and which forms a chamber to contain the exhaled breath within the apparatus (to be vented out appropriately), and (e) a sample collection mechanism. In one embodiment, the sample collection mechanism may comprise an elongated wick for absorbing the condensed droplets. In various embodiments, the collection mechanism could be a ring-shaped material located at the equilibrium radial location mentioned earlier. In other embodiments, the collection mechanism could be a wicking material, surface grooving/texturing, gravity-assisted, or external-flow assisted collection mechanism. In various embodiments, the apparatus may include a flow control system on the surface.

[0042] In various embodiments, the sampling apparatus described herein can provide low- cost monitoring system for point-of-care testing, with quick interfacing to analysis systems as well as environmental monitoring which are needed for re-opening public spaces, public safety, and protecting against the current and future pandemics.

[0043] In various embodiments, the design disclosed herein increases the efficiency of condensation from a humid air stream. In various embodiments, the combination of condensation and particle collection is unique from exhaled breath. Commercially- available systems generally use filters rather than solid plates and require one to dissolve the filter to test for viruses. In various embodiments, the fluidic handling on the substrate is unique and the integration of this exhaled breath collection system with testing techniques like fluorescent in situ hybridization has never been shown.

[0044] In various embodiments, the apparatus may be disposable. In various embodiments, the apparatus may be single-use. In various embodiments, the apparatus may be sized such that it may be held in a human hand ( e.g ., an adult hand).

[0045] Further aspects of the technology described herein will be brought out in the following portions of the specification, wherein the detailed description is for the purpose of fully disclosing preferred embodiments of the technology without placing limitations thereon.

[0046] In various embodiments, an apparatus is provided for collecting samples of condensed air moisture for viral load level diagnosis. Fig. 1A illustrates a flow diagram of an embodiment including a condensation apparatus 102 and a polymerase chain reaction (PCR) system 104. In various embodiments, humid air may be sampled from a patient’s breath or from environmental (e.g., room) air by the condensation apparatus 102, such as the apparatuses described herein. In various embodiments, the condensed moisture collected by the condensation apparatus 102 may be transported to the PCR system 104 for analysis of virus presence. In various embodiments, the PCR system may be any suitable commercially-available PCR system (e.g., qPCR).

[0047] Fig. IB illustrates a diagram showing a general method for continuous dropwise concentration (CDC) 112 and retained exhaled breath condensate (EBC) 114 for polymerase chain reaction (PCR) analysis of virus presence. In various embodiments, human exhaled breath may enter through a one-way flow restriction (e.g, one-way valve) to a continuous dropwise concentration 112 device. In various embodiments, the exhaled breath may exit the CDC device 112 via a one-way flow restriction. In various embodiments, the retained EBC 114 may be transported to a PCR system (e.g, rtPCR) for analysis of virus presence or an in-situ detection of viral load may be implemented.

[0048] In various embodiments, the condensation apparatus 102 described herein has been experimentally found to be more efficient than state-of-the art dehumidification systems with respect to collecting moisture from a human breath and/or the ambient air. Referring to Figs. 2A-2B, in various embodiments, the apparatus 200 directs a gaseous sample 250 containing moisture ( e.g ., a patient's breath or humid air) such that the gaseous sample impinges on a sample collection surface 202 (of a sample collection base 201) that is cooled down to a desired temperature that is lower than the dew point of the incoming air. In various embodiments, a patient may blow into a tube 204 (e.g., disposable tube) that is configured to direct and focus the patient's breath (i.e., the gaseous sample 250) as a jet of air toward the sample collection surface 202. In various embodiments, because the sample collection surface 202 (i.e., the surface facing the user) is cooled below the dew point of the incoming air, a breath figure (BF) spot appears quickly (e.g, almost instantaneously). In various embodiments, this BF spot may include one or more micron-sized water droplets 214a. In various embodiments, as the droplets grow to become larger droplets 214b, shearing effects of the jet passing over the droplets pushes the droplets radially outward to an equilibrium radial location. In various embodiments, these water droplets (containing the target, e.g, virus, bacteria, fungus, etc.) may be collected by using a collection material to absorb the condensed droplets 214a, 214b. In various embodiments, a ring shaped collection material may be placed at or near this equilibrium radial location. In various embodiments, a ring shaped collection material may be placed at a radius that is less than the equilibrium radial location to thereby collect additional droplets. In various embodiments, the liquid droplets can then be transported for analysis.

[0049] In various embodiments, the apparatus 200 is configured to function as a continuous dropwise condensation (CDC) based collector with a cooling device (not shown). In various embodiments, CDC combines hydrophobic surface properties with strong shear stress from an incoming jet of humid air or breath to continuously displace condensed droplets from a cold hydrophobic condensation zone to an annular hydrophilic collection zone from where it may be readily extracted. In various embodiments, humidified air flow is focused on the center of a cooled hydrophobic zone. In various embodiments, small drops condense and are forced by shear flow outward where they coalesce into larger drops that are driven outward by the flow and their larger size to a collection zone. In various embodiments, the collection zone may be a hydrophilic zone or a complex functionalized surface or hydrogel.

[0050] In various embodiments, the sample collection surface includes a sample collection material 210 disposed on the sample collection surface 202. In various embodiments, the sample collection material 210 may include a ring. In various embodiments, the ring of sample collection material 210 may be positioned on the sample collection surface 202 such that the center of the ring is aligned with the center of the tube 204. In various embodiments, the sample collection material 210 may be hydrophilic ( e.g ., have hydrophilic properties and/or be treated with a hydrophilic coating). In various embodiments, the sample collection material 210 may include a hydrogel. In various embodiments, the hydrogel may be functionalized with an enzyme attached thereto (described in more detail in Fig. 8). In various embodiments, at least a portion of the sample collection surface 202 may include hydrophobic surface properties, e.g., by applying a hydrophobic surface coating. For example, the area 205 within the sample collection material 210 may include a hydrophobic coating. In various embodiments, the sample collection surface 202 may include a substrate material 211 around the perimeter (e.g, circumference) of the sample collection material 210. In various embodiments, the substrate material 211 may include glass.

[0051] FIG. 2B shows an experiment demonstrating the ability of the apparatus to condense and collect droplets from humidified air (70-80% RH; 25°C) for approximately two minutes at a flow rate of 25 liters/min. In various embodiments, a 37°C, humid jet of exhaled air may be directed to the collecting disk. In various embodiments, the jet impinges upon the disk at the center of the subcooled, tuned, hydrophobic zone. In various embodiments, the impinged jet expands in the radial direction as it approaches the surface. In various embodiments, the temperature of the substrate may be maintained below the dew point of breath, typically approximately 5°C, triggering immediate condensation on the central surface. In various embodiments, as the newly condensed droplets grow (R ~ t0.4), the shear force from the radial flow stream may overcome the contact force and drives them out of the central condensing zone. In various embodiments, as drops move outward, the droplets coalesce, collecting other droplets along the way, increasing their surface area and drag. In various embodiments, as the radial flow field expands and slows, the droplets, enlarged via coalescence, present a larger form (diameter) and surface area to the flow enabling their continued outward migration to the hydrophilic collection zone. In various embodiments, the process clears the central surface permitting continuous dropwise condensation or CDC. In various embodiments, the volume of condensed water grows faster than linearly with time, V ~ tl .2.

[0052] In various embodiments, parameters that control the collection rates and condensate distribution may include: (1) substrate temperature; (2) surface properties; (3) incoming jet exit speed; (4) jet diameter; (5) jet standoff distance; and (6) zone radii on the disk. In various embodiments, manipulation of these parameters permit the rate of condensate volume accumulation to be optimized. In various embodiments, a physiologically relevant jet of exhalate at 60 LPM may result in about 1.3 mL/min of collected condensate.

[0053] In various embodiments, other EB collection techniques transition from dropwise to film condensation over time as water is accumulated, which dramatically reduces the mass transfer coefficient and the rate of water accumulation on the surface. In various embodiments, the collection apparatus presented herein maintains dropwise condensation throughout the collection process thereby maximizing the rate of condensation. In various embodiments, existence of multiple stagnation points at the center of the disk, as well as on already-formed droplets throughout the collection area, may enhance the liquid entrapment of aerosolized particles with sizes from submicron to multiple microns.

[0054] Fig. 3A illustrates an exhaled droplet condensation collection apparatus 300. In various embodiments, the apparatus 300 comprises (a) a sample collection surface 302 (of a sample collection base 301), at least a portion ( e.g ., area 305) of which may be optionally treated with hydrophobic material, (b) a tube 304 configured to direct a patient's exhaled breath thereby impinging the focused jet of breath on the collection surface, (c) a cooling device 306 that cools the collection surface with sufficient cooling capacity to form condensation, (d) a cover 308 or enclosure coupled to the tube 304 and which forms a chamber to contain the exhaled breath within the apparatus (to be vented out appropriately), and (e) a sample collection material 310. In various embodiments, the sample collection base 301 and the cover 308 collectively form a housing that is substantially enclosed (e.g., except for the ports and vents). In various embodiments, the apparatus 300 may have a volume of about 3 inches x 3 inches x 2 inches.

[0055] In the embodiment illustrated in Fig. 3A, the cover 308 fits over the sample collection surface 302 to form a sample collection chamber 320 having a substantially- enclosed volume. In various embodiments, the cover 308 has at least one inlet port 309. In various embodiments, where the tube 304 is disposable, the distal end of the tube 304 may be inserted to provide fluid communication between a proximal end of the tube 304 and the substantially-enclosed chamber. In various embodiments, the tube 304 may be integral with the cover such that the lumen of the tube 304 is aligned with the inlet port 309 of the cover 308. In various embodiments, the tube 304 extends through the port such that a distal opening of the tube 304 is in fluid communication with the sample collection chamber 320. Preferably, the cover has a single inlet port 309. In various embodiments, the at least one inlet port 309 may be position substantially at the center of the cover. In various embodiments, the tube may include a one-way valve 316 configured to allow the flow of the gaseous sample in a single direction from a user’s mouth into the sample collection chamber 320 thereby preventing backflow. In various embodiments, the tube 304 may include a cover 313 over the end that is configured to receive the user’s mouth. In various embodiments, the cover 313 may be a tamper-evident cap configured to indicate if the apparatus has been used. In various embodiments, the cover 313 may be removed prior to use.

[0056] In various embodiments, the tube 304 may have a substantially smaller diameter than the mouthpiece diameter. In various embodiments, the tube 304 may be surrounded by a thick wall that extends the overall diameter to that of the mouthpiece. In various embodiments, the tube 304 is illustrated as small diameter orifice interior to a thick-walled mouthpiece. In various embodiments, the diameter of the tube 304 can vary in relation to the mouthpiece diameter. In various embodiments, while other positions are possible, preferably the tube 304 is centered in relation to the center of the substrate and aligned along a vertical axis that is orthogonal to the surface of the substrate.

[0057] In various embodiments, a sample collection material 310 may be disposed ( e.g ., adhered) to the sample collection surface 302. In various embodiments, a substrate material on the sample collection surface 302 may include glass, polymer, polytetrafluoroethylene, and/or other materials. In various embodiments, the sample collection surface 302 may be modified by treating it with a hydrophobic material and preferably with high contact angles. In various embodiments, the sample collection material 310 may be a ring. In various embodiments, the sample collection material 310 may include a hydrogel configured to absorb condensed moisture from the gaseous sample. In various embodiments, the hydrogel may be functionalized with one or more molecule, such as an enzyme (e.g., ACE2), to which a target (e.g, SARS-CoV-2, bacteria, fungi, pathogen, etc.) may bind. In various embodiments, the inlet port 309 may be positioned relative to the collection material 310 (e.g, hydrogel) such that the droplets are absorbed by the collection material. In other embodiments, the inlet port 309 may be positioned at the center of the sample collection material 310, such as in the embodiment where the sample collection material 310 is ring-shaped. [0058] In various embodiments, the sample collection surface 302 may include a hydrophilic or a functionalized hydrogel collection ring. For example, to capture intact functional virus particles from the EB, the condensing droplets can be collected by a ring of ACE2 functionalized adhesive hydrogel placed at an optimized location as shown.

[0059] In various embodiments, the sides of the cover 308 may be transparent so that the patient/technician can monitor droplet collection. In various embodiments, a temperature indicator tape/dye may be provided to tell the technician/user that the plate is ready for collection (not shown). In various embodiments, the apparatus preferably is a single-use device that would be packaged and distributed in a sterile blister pack.

[0060] In various embodiments, the base 301 (including the sample collection surface 302) of the sample collection chamber 320 may be cooled via a cooling device 306. In various embodiments, the cooling device 306 may be a thermoelectric device beneath the base 301 of the sample collection chamber 320. In various embodiments, the cooling device 306 may be a Peltier plate thermally coupled ( e.g ., thermally pasted) to the underside of the base 301 of the sample collection chamber 320. Other suitable methods known in the art could be used as well, provided that the cooling capacity is sufficient to form condensation from the gaseous sample. In various embodiments, any suitable cooled solid, liquid, gas, and/or combination of phases could be used as a cooling device. For example, ice (FhO), dry (CO2) ice, or liquid nitrogen could be placed in contact with the base 301 to thereby cool the base such that condensation will form as the gaseous sample is received in the sample collection chamber 320.

[0061] In various embodiments, the sample collection material 310 may include a ring- shaped material. In various embodiments, at least a portion (e.g., all) of the sample collection material 310 may be located at the equilibrium radial location on the sample collection surface 302. In various embodiments, the collection material 310 may include a ring-shaped material located at the equilibrium radial location as described above. In various embodiments, the sample collection material 310 may be integrated into (e.g, adhered to) a card or chip such that the card or chip collects condensed water droplets and can be removed once a suitable amount of condensed water droplets are collected. In various embodiments, the sample collection material may include any suitable shape (e.g, flat circular area, flat square area, square perimeter, ovular parameter, annular shape, or any other suitable symmetric or asymmetric shape such that the sample collection material collects condensed moisture from the sample collection surface). [0062] In various embodiments, the sample collection surface 302 may be treated with receptors to one or more portions of a virus ( e.g ., virus coat protein, virus spike protein, etc.), such as for the COVID-19 virus. In various embodiments, after exposure to exhaled air or ambient air, the sample collection surface 302 may be placed into a processor where the unoccupied receptors can be illuminated fluorescently with known immunohistochemical methods. In various embodiments, the virus may be labelled with anti-virus (e.g., anti-COVID-19) antibodies, producing a direct readout of virus titer. In various embodiments, a direct testing “lab-on-a-chip” system may be provided.

[0063] In various embodiments, the apparatus may be configured to bind one or more antigen, for example, bacteria, fungus, single-celled organisms, etc. In various embodiments, the apparatus may be configured to bind to any suitable target that may be exhaled from the lung(s) via aerosols.

[0064] In various embodiments, the cover 308 may include one or more vents 312. In various embodiments, the one or more vents 312 may be disposed such that the vents are disposed about the circumference of the cover 308. In various embodiments, the one or more vents 312 may be equally-spaced around the cover 308. In various embodiments, the one or more vents 312 may be rectangular cutouts. In various embodiments, the one or more vents 312 may be circular cutouts. In various embodiments, the one or more vents 312 may be square cutouts. In various embodiments, the one or more vents 312 may be oval cutouts. In various embodiments, the one or more vents 312 may have any suitable shape to provide for outflow of the gaseous sample. In various embodiments, the one or more vents 312 may include a one-way valve.

[0065] In various embodiments, the cover 308 may include a swab port 318. In various embodiments, the swab port may have a diameter that is larger than a diameter of the one or more vents 312. In various embodiments, the diameter of the swab port 318 may be sized such that a swab can be inserted into the swab port 318. In various embodiments, the swab port 318 may include any suitable shape, such as, for example, a circular cutout. In various embodiments, the swab port 318 may include a removable cover. In various embodiments, as shown in Fig. 3A, the cover 308 may have a constant area (i.e., constant diameter) along a longitudinal axis defined by the lumen of the tube 304. For example, the cylindrical shape of the cover 308 has a constant diameter from the inlet port 309 to the sample collection surface 302. In various embodiments, the inlet port 309 has a first diameter and the cover 308 has a second diameter that is larger than the first diameter in a step-wise manner. In various embodiments, the cover 308 may be transparent. In various embodiments, the base 301 (and the sample collection surface 302) may be transparent. [0066] In various embodiments, a diameter of the tube 304 may be about 1 mm to about 5 mm. In various embodiments, the diameter of the tube 304 may be about 2 mm to about 3 mm. In various embodiments, a height between the distal end of the tube 304 and the sample collection surface 302 may be about 1mm to about 200mm. In various embodiments, the height between the distal end of the tube 304 and the sample collection surface 302 may be about 1mm to about 100mm. In various embodiments, the height between the distal end of the tube 304 and the sample collection surface 302 may be about 1.5 mm to 52.5 mm. In various embodiments, the flow rate of the gaseous sample ( e.g ., humid air) through the tube 304 may be about 0.5 liter per minute (LPM) to about 5 LPM. In various embodiments, the flow rate of the gaseous sample (e.g., humid air) through the tube 304 may be about 1 LPM to about 3 LPM. In various embodiments, a surface temperature of the sample collection surface 302 may be about 0°C to about 22°C. In various embodiments, a surface temperature of the sample collection surface 302 may be about 22°C to a temperature of 5°C.

[0067] Fig. 3B illustrates an embodiment of an exhaled droplet condensation collection apparatus 300. The exhaled droplet condensation collection apparatus 300 is similar to the exhaled droplet condensation collection apparatus 300 of Fig. 3 A, with the cover 308 of the apparatus 300 having a shape of an Erlenmeyer flask and the tube 304 being integral with the cover 308. In various embodiments, as shown in Fig. 3B, the cover 308 may have a varying area (i.e., varying diameter) along a longitudinal axis defined by the lumen of the tube 304. For example, the Erlenmeyer flask shape has an increasing diameter from the inlet port 309 to the sample collection surface 302. In various embodiments, the inlet port 309 has a first diameter and the cover 308 has a constantly increasing (e.g, linearly increasing) diameter.

[0068] It will be appreciated that the presented technology can be used in connection with detection of a variety of different viruses, bacteria, etc. and/or antibodies for diseases. [0069] In various embodiments, the apparatus described herein may be used in any suitable type of airflow for sample collection, with two main applications related to current pandemic being human sampling and using natural ambient airflow to provide long term monitoring of important enclosed environments such as hospital, offices, schools, airport terminals, etc. [0070] Figs. 4A-4B illustrate a comparison of an exhaled breath collection system (Fig. 4A) and an embodiment of an apparatus according to the present disclosure (Fig. 4B). In particular, known exhaled breath systems (shown in Fig. 4A) may have a height of about 5 feet while the apparatuses of the present disclosure (shown in Fig. 4B) have a height of about 2 inches, significantly smaller than known EB systems.

[0071] Fig. 5 illustrates a system diagram of baseline use of an exhaled breath collection system. In various embodiments, the apparatus may be compatible with existing and emerging point-of-care (POC) diagnostic test systems. Fig. 5 illustrates a basic use of the EB apparatus in combination with a commercially-available rtPCR-based diagnostic testing system. In various embodiments, the collected sample may be extracted from the collection zone and transported ( e.g ., shipped) to a location where diagnostic testing (e.g, rtPCR) occurs. In various embodiments, where the sample collection surface is on a removable and/or disposable chip, the chip may be removed from the EB apparatus and transported to a location where diagnostic testing (e.g, rtPCR) occurs.

[0072] In various embodiments, the apparatuses and systems described herein may be integrated into a fluorescence in situ hybridization (FISH) test to allow for point of care or at-home diagnostics for a virus (e.g., SARS-CoV-2).

[0073] Fig. 6 illustrates a system diagram illustrating a point-of-care system with fluorescence in situ hybridization (FISH) developer and scanning. In particular, FIG. 6 illustrates use of the EB apparatus as the front end for a sensitive, rapid POC diagnostic test using a modified version of a method for rapid single molecule RNA FISH (e.g, TurboFISH). In various embodiments, unlike nasal swabbing, the EB apparatus provides for CDC-based sample collection on a 2-D plate thereby enabling a large range of on-chip detection opportunities via surface functionalization combined with techniques such as fluorescence in situ hybridization, ELISA or immunohistochemistry. In various embodiments, single RNA detection using FISH can be performed on virus particles captured on the collection plate in an ACE2 seeded hydrogel. In various embodiments, the plate can also be modified to port the sample to a separate microfluidic platform for off- chip local diagnostics as they are developed for viral testing.

[0074] In various embodiments, for COVID-19 detection, FISH may be an appropriate on board detection method. In various embodiments, FISH may be used for single RNA detection with high specificity and sensitivity coupled with high signal amplification. In various embodiments, FISH may be used to detect viruses, and may be automated for on- chip detection and throughput.

[0075] In various embodiments, it will be appreciated that the collection/detection system can provide not only a positive or negative diagnosis, but it can also quantify the amount of virus in the exhaled breath. In various embodiments, information related to the viral load may essential for the health care provider to have an informed view of the state of the disease and the current level of contagiousness of the patient.

[0076] In various embodiments, because the EB apparatus allows for a short time from sample to result, information can quickly flow to a physician, and guidance can be provided back to the patient during the same visit. In various embodiments, the EB apparatus enables full clinical integration of point-of-care testing and timely exchange of vital information. In various embodiments, test results (combined with physiological interpretations) allow physicians to plan prospectively rather than retrospectively. In various embodiments, for cases involving COVID-19, connectivity of point-of-care test results to distant physicians is essential to facilitate diagnosis, enhance therapy, and prevent errors. In various embodiments, there may be minimal ( e.g ., none) delay that could cause someone to quarantine unnecessarily, or worse, to infect others while waiting for a test result.

[0077] As shown in Fig. 6, a total process time from sample collection to result may be about 12 minutes and 25 seconds. In various embodiments, the total process time from sample collection to result may be less than about 30 minutes. In various embodiments, the total process time from sample collection to result may be less than about 20 minutes. In various embodiments, the total process time from sample collection to result may be less than about 15 minutes. In various embodiments, the total process time from sample collection to result may be about 12.5 minutes.

[0078] In various embodiments, a user may provide a breath sample into a tube of the exhaled breath collection apparatus and moisture from the breath sample may be condensed on the sample collection surface. In various embodiments, the user may be instructed to blow into the tube for a predetermined amount of time suitable to condense enough moisture for diagnostic testing. In various embodiments, the predetermined amount of time may be about 2 minutes. In various embodiments, the predetermined amount of time may be about 1.5 minutes. In various embodiments, the predetermined amount of time may be about 1 minute and 20 seconds. In various embodiments, the predetermined amount of time may be about 1 minute. In various embodiments, the predetermined amount of time may be less than 1 minute.

[0079] In various embodiments, the collected sample may be transferred to a FISH developer process ( e.g ., TurboFISH developer). In various embodiments, where the sample collection material is disposed (e.g., adhered) to a chip, the chip may be removed from the EB collection apparatus for on-chip processing. In various embodiments, the sample may be processed with methanol. In various embodiments, one or more probes may be attached to a target (e.g, RNA) or non-target within the sample. In various embodiments, the sample may be washed one or more times to isolate the target. In various embodiments, a FISH developer may include a visible light cross-linking process. In various embodiments, a FISH developer may include one or more fluid handlers selected from: methanol fix,

FISH probes, and wash steps. In various embodiments, the sample (after the FISH developer process) may be transferred to a scanner. In various embodiments, each transfer process may take about 30 seconds. In various embodiments, the scanner includes one or more LEDs configured to excite the FISH probes. In various embodiments, the scanner includes a digital camera. In various embodiments, the scanner includes an image processor configured to receive images from the digital camera and process the images based on the excitation of the FISH probes. In various embodiments, transferring the chip (e.g, a disk) after the FISH developer process includes affixing the chip to a rotational platform coupled to a motor. In various embodiments, at least a portion of the rotational platform is transparent. In various embodiments, the scanner includes a 40x long working distance lens aligned with the sample collection material (e.g, hydrogel functionalized with ACE2) ring.

[0080] Fig. 7 illustrates a flow diagram of a system workflow for sample collection and analysis. Furthermore, as aspect of the technology is to detect functional virus rather than just virus RNA (as is done with rtPCR based methods). In various embodiments, a baseline system workflow may include a patient check-in process (~2 minutes) where a patient is logged-in, a sample collection apparatus is labelled, and a shipping tube is labelled. In various embodiments, the baseline system workflow includes a test administration process (~2 minutes) where the sample collection apparatus is unpacked, the sample collection apparatus is placed on a cooling plate (e.g, thermoelectric plate), the patient is instructed on how to use the collector, seals are removed from the tube and/or vents of the collector, and sample collection is monitored (e.g, by a healthcare provider). In various embodiments, the baseline system workflow includes a sample retrieval step (~1 minute) where the port cover on the sample collection apparatus is removed, the exhaled breath condensate is extracted from the chip ( e.g ., a disk), and the extracted fluid is placed in the shipping tube. In various embodiments, the shipping tube is transported to a location to perform diagnostics on the sample, as described above.

[0081] In various embodiments, a point-of-care system workflow may include a run developer process (~1 minute of tech time and ~9 minutes of processing) where the collection chip (e.g., a disk) is placed into a FISH develop, fluid levels are checked, and the FISH developer process is started (as described above). In various embodiments, the point- of-care system workflow includes a run scan process (~1 minute) where the collection chip (after processing in the FISH developer) is transferred to a FISH scanner, the collection chip is scanned, the results are logged, and the collection chip is stored for a suitable amount of time (e.g, as required by regulatory laws).

[0082] In various embodiments, because it is time consuming to examine the entire volume of fluid for virus, the collected virus may be concentrated in a small, easily interrogatable region of the sample collection surface. In various embodiments, to capture intact functional virus particles, in one embodiment the condensing droplets may be collected by a ring of ACE2 functionalized adhesive hydrogel placed at an optimized location.

[0083] Fig. 8A illustrates a chemical reaction for conjugation of ACE2 to hydrogel 1HNMR spectra of hydrogel (10% and 20% (w/v)) formed at varying visible light exposure times, including 0, 1, 2, and 4 minutes. In various embodiments, fabrication of ACE2 functionalized adhesive hydrogel includes two independent chemical processes: covalent conjugation of gelatin macromolecules with methacrylic anhydride followed by the functionalization of the modified gelatin with an enzyme, for example, angiotensin converting enzyme 2 (ACE2). In various embodiments, incorporation of acrylic moieties imports the adhesive property upon photocrosslinking with visible light. In various embodiments, methacrylic anhydride may be added to the gelatin solution to produce a modified gelatin. In various embodiments, to produce an enzyme-enriched (e.g, ACE2) ring, an enzyme (e.g, ACE2) may be covalently bonded to the backbone of polymer through linkages (e.g, amide or imine). In various embodiments, other suitable linkages as are known in the art may be used. In various embodiments, reactions may involve the use of a linker molecule such as glutaric dialdehyde, which efficiently binds the ACE2 to the hydrogel backbone. In various embodiments, the target (e.g, virus, bacteria, fungus, etc.) may be immobilized directly on the glass surface through standard glass functionalization methods (adhered biotin, etc). In various embodiments, when there is any variation in the height of the bound virus above the glass, the scanning time may be increased. In various embodiments, effective scanning may be performed when all bound, labelled virus are no more than 1 micron away from the plate.

[0084] Fig. 8B illustrates a coating of device ( e.g ., a sample collection chip) with ACE2- hydrogel. In various embodiments, to form bioadhesive hydrogels, a predetermined concentration of photoinitiator is added to the modified gelatin solution and exposed to visible light for one minute to photopolymerize the adhesive hydrogel on the surface of the collection plate. In various embodiments, following the attachment of ACE2 to the hydrogel prepolymer, an aqueous solution of ACE2 functionalized hydrogel prepolymer may be mixed with a photoinitiator. In various embodiments, the solution may be used to form a thin layer with thickness of 1-2 pm on the surface of the device by spin coating the hydrogel solution, followed by a short visible light photocrosslinking step (~1 min.). In various embodiments, an additional layer of hydrogel without ACE2 (50-100 pm thickness) may be photopolymerized on the top of ACE2-hydrogel layer to collect the liquid containing virus. In various embodiments, once the virus diffuses inside the hydrogel matrix, the hydrogel may be further crosslinked by exposing to visible light for another 30 seconds to enhance the degree of the crosslinking inside the hydrogel matrix and therefore prevent the diffusion of RNA out of the hydrogel network.

[0085] Figs. 9A-9E illustrate a method for performing FISH using condensed breath. In particular, Fig. 9A illustrates a top view of a sample collection surface 302 that may be chilled to a suitable temperature (e.g., a cold substrate) to thereby cause condensation of moisture contained within human breath and collection of virus particles. In various embodiments, the material of the sample collection base 301 and/or sample collection surface 302 may be any suitable material having thermal properties (e.g, heat capacity, conductivity, etc.) and/or thickness configured to cool incoming breath such that condensed moisture forms on the sample collection surface 302. Fig. 9B illustrates a side, cross- sectional view of the apparatus 300 where a user has provided an inflow of warm humid breath through the tube 304 (e.g, a straw) with virus particles (represented as black dots) entrained in the flow of the breath. Fig. 9C illustrates a side, cross-sectional view of the apparatus 300 where humidity in the breath (containing the virus particles) condenses on the sample collection surface 302 and the shear from the incoming breath drives growing liquid droplets outward from the center of the forming ring. Fig. 9D illustrates a side, cross-sectional view of the apparatus 300 where the virus particles in the breath impact the sample collection surface 302 and stick to condensed water droplets on the sample collection surface 302. Fig. 9D illustrates a side, cross-sectional view of the apparatus 300 where successive breaths increase the volume of condensed liquid on the sample collection surface 302 and the number of virus particles captured in the liquid droplets.

[0086] Figs. 10A-10E illustrate a method for performing rtPCR using condensed breath in an apparatus 300 using a swab 330 for moisture collection. In particular, Fig. 10A illustrates a top view of a sample collection surface 302 that may be chilled to a suitable temperature to thereby cause condensation of moisture contained within human breath and collection of virus particles. The sample collection base 301 and/or sample collection surface 302 may be made from a polymer, e.g ., high density polyethylene (HDPE).

Fig. 10B illustrates a side, cross-sectional view of the apparatus 300 where a user has provided an inflow of warm humid breath through the tube 304 (e.g, a straw) with virus particles (represented as black dots) entrained in the flow of the breath. As shown in Fig. 10B, the sample collection base 201 includes a thickness H that has suitable thermal properties (e.g, conductivity, heat capacity, etc.) to condense moisture from human breath when in contact with a cooling device, such as the cool side of a thermoelectric plate.

Fig. IOC illustrates a side, cross-sectional view of the apparatus 300 where humidity in the breath (containing the virus particles) condenses on the sample collection surface 302 and the shear from the incoming breath drives growing liquid droplets outward from the center of the forming ring. Fig. 10D illustrates a side, cross-sectional view of the apparatus 300 where the swab port 318 is opened and a swab 330 is inserted therethrough to collect condensed moisture from the sample collection surface 302. Fig. 10E illustrates a side, cross-sectional view of the apparatus 300 where, after the condensed moisture has been collected, the tip of the swab 330 is broken off, placed in a transport medium in a container 340, and sent for rtPCR testing. In various embodiments, the procedure may be substantially similar to the procedure for processing nasal swabs. In various embodiments, the apparatus 300 may be disposed of after use.

[0087] Figs. 11A-11F illustrate a method for performing rtPCR using condensed breath in an apparatus 1100 having microfluidic channels 1160. In particular, Fig. 11A illustrates a top view of a sample collection surface 1102 of a sample collection base 1101 (collectively, 1101’ and 1101”) having one or more microfluidic channels 1160 configured to act as gutters for condensed moisture. The sample collection surface 1102 (of the sample collection base 1101) is configured to be cooled to a temperature to condense moisture within human breath. The microfluidic channels 1160 include a central ring 1160a and an outflow channel 1160b configured to collect condensed moisture. The central ring 1160a and the outflow channel 1160b each have a width w and the central ring 1160a has an outer radius Rout. Fig. 11B illustrates a side, cross-sectional view of the apparatus 1100 where a user has provided an inflow of warm humid breath through the tube 1104 ( e.g ., a straw) and into the sample collection chamber defined by the cover 1108 and the sample collection base 1101. In various embodiments, the sample collection base is made of a substrate layer 1101’ (e.g., a glass substrate) and an upper layer 1101” including a treated coating (e.g., hydrophobic coating or polymeric film). In various embodiments, the substrate layer 1101’ may include a hydrophilic coating on at least a portion of the substrate layer 1101’ (e.g, just the exposed portions or the entire layer). In various embodiments, the apparatus 1100 further includes an outlet 1118 for collection of the condensed moisture. Fig. 11C illustrates a side, cross-sectional view of the apparatus 1100 where humidity in the breath (containing the virus particles) condenses on the sample collection surface 1102 and the shear from the incoming breath drives growing liquid droplets outward from the center of the forming ring thus collecting in the microfluidic channels 1160 as condensation 1162. Fig. 11D illustrates a side, cross-sectional view of the apparatus 1100 where a transport medium is added to the collected condensation 1162 in the microfluidic channels 1160. In various embodiments, the transport medium may be added via the tube 1104 to thereby rinse the sample collection surface 1102 and fill the microfluidic channels 1160. In various embodiments, the transport media may be a solid (e.g, a coating or powder) that may be dissolved upon contact with condensed human breath. Fig. HE illustrates a side, cross- sectional view of the apparatus 1100 where a container 1140 (e.g, centrifuge vial) is placed over the outlet 1118 and the apparatus 1100 is tilted sideways to thereby cause the condensation (optionally, with added transport media) to flow into the container 1140. In various embodiments, the container 1140 may be configured to removably couple to the outlet 1118 via, for example, a screw-on, snap-on, or geometric fitting (e.g, taper). Fig. 11F illustrates the container 1140 after being sealed with a cap (e.g, a snap-on cap with tamper-evident seal) and ready for transport to a lab for diagnostic testing.

[0088] Figs. 12A-12D illustrate a method for performing FISH using condensed breath in an apparatus 1200 having microfluidic channels 1160. In particular, Fig. 12A illustrates a top view of a sample collection surface 1202 of a sample collection base 1201 (collectively, 1201’ and 1201”) having one or more microfluidic channels 1260. The sample collection surface 1202 (of the sample collection base 1201) is configured to be cooled to a temperature to condense moisture within human breath. The microfluidic channels 1260 include a central area 1260a, two or more spokes 1160b, and an outer ring 1260c, each configured to collect condensed moisture. In various embodiments, the central area 1260 is configured to allow light to pass through, e.g ., for microscope access during FISH processing. The outer ring 1260c has a width w and each spoke 1260b has a width w spoke. The central area 1260a has an inner radius, Rm , and the outer ring has an outer radius, Rout. Fig. 12B illustrates a side, cross-sectional view of the apparatus 1200 where a user has provided an inflow of warm humid breath through the tube 1204 (e.g, a straw) and into the sample collection chamber defined by the cover 1208 and the sample collection base 1201. In various embodiments, the sample collection base is made of a substrate layer 1201’ (e.g, a glass substrate) and an upper layer 1201” including a treated coating (e.g., hydrophobic coating). Fig. 12C illustrates a top view of the apparatus 1200 where humidity in the breath (containing the virus particles) condenses on the sample collection surface 1202 and collects in the microfluidic channel 1260 as condensation 1262. Fig. 12D illustrates a side, cross-sectional view of the apparatus 1200 where the condensation 1262 is disposed within the microfluidic channels. In various embodiments, one or more process chemicals may be added to the collected condensation 1262 in the microfluidic channel 1260 via the tube 1204 to thereby prepare the condensation 1262 for FISH processing.

[0089] In various embodiments, the width of the microfluidic channels may be about 1/16th to about 3/16th of an inch.

[0090] In various embodiments, the thickness, t, of the sample collection base and the material of use may be chosen such that the temperature of the bottom surface can cool quickly during preparation and remain cool during use. In various embodiments, it may be desirable that the bottom surface cool from room temperature at 20°C to 5°C in less than 10 seconds after being placed on a cold surface (e.g, chilled metal surface) held at between 0°C and 1°C. In various embodiments, a ratio of thermal conductivity to length greater than k/t > 300 W/m^A2K may be used. In various embodiments, this can be achieved with HDPE at a thickness of about 1/16 in (1.6mm). In various embodiments, the substrate material may be selected such that it maintains a cool temperature after it is removed from the cold substrate. In various embodiments, the material may be selected such that it has as large a specific heat, c, as possible. In various embodiments, it may be preferable for the specific heat to be greater than c > 1.5 kJ/kg/K. As an example, HDPE has a specific heat of c = 1.9kJ/kg/K. In various embodiments, the material may be chosen to maximize the rate of heat transfer to the condensing water vapor. In various embodiments, the effusivity of the surface may be maximized. In various embodiments, the effusivity may be defined as the square root of the density times the specific heat times the thermal conductivity, (rho c k) ^L 1/2. In various embodiments, an effusivity > 25 J/m^A2/K/s^Al/2 may be desirable. As an example, HDPE has an effusivity of 29 J/m^A2/K/s^Al/2.

[0091] In various embodiments, the design of the collection apparatus enables ease of use for the operator in addition to capability of incorporating the collection apparatus into a “touchless” automated system to retrieve the sample from the collector and deliver it to the detection system ( e.g ., PCR) without any need of human exposure, either using robotic fluid handling systems or fluidic system.

[0092] In various embodiments, the apparatus may be used with post-processing of biological samples (e.g., cell lysis) on the sample collection surface in order reduce losses during transfer of the collected sample.

[0093] In various embodiments, other than a Peltier plate, many other suitable passive and active cooling system can be used to set the sample collection surface temperature to the desired value to condense moisture from an incoming stream of human breath.

[0094] In various embodiments, the tube (e.g, straw) may be positioned at a distance of about 3 to 5 times the outlet diameter of the tube from the cooled sample collection surface. In various embodiments, the diameter of the outer wall of the device may be at least lOx of a diameter of the tube (e.g, inner diameter or outer diameter).

[0095] Fig. 13 illustrates an experimental setup used for condensate extraction from a humid air jet in accordance with an embodiment of the present disclosure.

[0096] Example 1 (Experimental Procedure)

[0097] Fig. 14 shows the BF spots as photographed from a side view using a regular camera. In various embodiments, BF spots may appear cloudy because micron-sized droplets scatter light in all directions. This enables easier visualization of the spots and better quantification of BF spot behavior. In various embodiments, height and Reynolds number may have minimal effects on the extent to which BF spots expand. In the tested range, BF spots varied from one to ten tube diameters. [0098] Fig. 15 shows infrared (IR) camera snap shots of selected cases at different surface temperatures. As shown in Fig. 15, at the lower the surface temperature, the higher water droplet are collected. Additionally, for a given surface temperature, the water collected increases with time. As droplets grow, the shearing effects of the jet may push the droplets radially outward to an equilibrium location at which they could be collected. In various embodiments, the radial equilibrium location may be controlled by using different surface wettability coatings. In various embodiments, the surface wettability coating may include a hydrophobic silicon polymer ( e.g ., hydroxy -terminated polydimethylsiloxane).

[0099] Example 4 (Numerical Example)

[0100] In various embodiments, the sample collection apparatus may be design such that a person with a limited lung capacity could exhale sufficient water vapor to collect a sample in under a minute. For example, consider a person exhaling (temperature 7) = 35°C and relative humidity 0_j = 100%) through a tube of diameter (D = 2mm) onto a surface that is cooled to a temperature of ( T_s = 5°C). Based on experimental measurements, the mass transfer coefficient was found to be (h_mA = 0.0166 g/s) if the breath flow rate is (Q = 3LPM). The mass flow rate for these settings is: m = h_mA (w_¥ - w₅) = 1.32 x 10³ (1.26 x 10-⁵)(0.0353 - 0.0054) * 5 x 10^®g/s [0101] If one exhales for 1 minute, around 0.03cc of liquid may be collected. Referring to Fig. 13, in various embodiments, human expiratory flow rates can reach around 300 LPM. In various embodiments, a condensation rate of 2mL/min is sufficient for diagnostic testing. [0102] As described above, this disclosure provides for a highly efficient sample collection system, based on a novel condensation approach, that can convert humidified air from patient exhalation or directly from the local environment into a highly concentrated sample from which the virus titer can be extracted.

[0103] In various embodiments, the system is more efficient, and potentially up to lOOx more efficient, at collection than any other Exhaled Breath Condensation (EBC) technique currently available giving us the potential to make rapid point of care collection and diagnosis more reliable. While existing EBC techniques can take 30 minutes to an hour of breathing through a tube in order to collect sufficient samples, in various embodiments, the presented technology can provide results in less than a minute.

[0104] In various embodiments, the presented collection system can easily and seamlessly be integrated into number of different detection systems including PCR and possibly future microfluidic platforms in a safe, clean and reusable way without cross-contamination between uses.

[0105] Fig. 17 shows a configuration of a jet exiting a nozzle and impinging on a surface in a quiescent ambience. In various embodiments, the jet may be submerged or non- submerged depending on whether the ambience is of similar density to the jet fluid. In various embodiments, the jet can be divided into three regions; (1) Free jet region, (2) stagnation/impingement region, and (3) wall jet region. In various embodiments, in the free jet region, flow, thermal, and species fields may not be affected by the solid wall downstream. Therefore, the variation of the state variables of this region can be matched with a free unbounded jet. In various embodiments, the free jet can be divided further into developing and developed regions, similar to that in pipe flow. In various embodiments, the developing region is characterized by a potential core that maintains the velocity profile of the nozzle exit. In various embodiments, momentum, heat, and mass are exchanged with the surroundings. In various embodiments, state variable variation may have a smooth transition. In various embodiments, this region may extend to almost five nozzle diameters. In various embodiments, in the developed region, the potential core may vanish, and the center line magnitude of state variables may start changing. In various embodiments, in the stagnation region, the speed of the jet may drop to zero at the center of the impingement area and the pressure may build up to a maximum of (pv²/ 2). In various embodiments, heat and mass transfer rates in the stagnation area may be significantly higher. In various embodiments, in the wall jet region, a regular boundary layer problem may be solved with prior knowledge of the boundary conditions. In various embodiments, several empirical correlations may be obtained under different conditions of single-phase heating or cooling applications. In various embodiments, a heat/mass transfer analogy may be utilized as well for drying processes. In various embodiments, in nucleate boiling, jet impingement may be observed and models may be used to estimate the boiling curve for such cases.

[0106] Many cases in single-phase jet impingement include a condition of a turbulent nozzle exit, which assumes a uniform velocity profile at the nozzle exit. Some reviewers have addressed most of physical phenomena and empirical correlations of gas jet impingement on solid surfaces, while other reviewers have focused on liquid jet impingement heat transfer. A major difference between gas and liquid jet impingement is the existence of a hydraulic jump in the latter case. In various embodiments, heat transfer by jet impingement may be affected by several parameters. In various embodiments, nozzle configuration, nozzle diameter, nozzle-to-surface spacing, jet velocity, and the mismatch between jet and ambient temperature and concentration may affect the impingement.

[0107] In various embodiments, the nozzle geometrical configuration may include shapes, such as Square-edged orifices, standard-edged orifices, and sharp-edged orifices. In various embodiments, the nozzle may include round nozzles, e.g ., round nozzles with arrays of triangular tabs. In various embodiments, the nozzles may be round, square, and/or rectangular nozzles. In various embodiments, the nozzle configurations may be an important factor for enhancing the turbulent mixing of the jet. In various embodiments, turbulent mixing acts to enhance the heat transfer significantly. In various embodiments, comparison with fully-developed pipe jet impingement may show improvement as high as 55% in stagnation region heat transfer. In various embodiments, improvements as high as 75% may be achieved by replacing contoured nozzles with orifice nozzles. In various embodiments, round nozzles may produce the least pressure drops compared to square or rectangular nozzles. In various embodiments, length scales may be normalized by the nozzle diameter (or radius). In various embodiments, length scales may be normalized in the free jet region, the stagnation region, and/or wall jet regions. In various embodiments, the height-to-diameter ratio (H/D) may affect some variables. In various embodiments, for a uniform nozzle exit velocity, the Nusselt number radial distribution may function in two ways. For heights higher than five nozzle diameters, the distribution may be characterized by a bell-shaped curve for which Nu monotonically drops from the stagnation point outward. For lower heights, there exists a secondary peak in Nusselt number distribution that may exceed the stagnation point peak. In various embodiments, a height value of five diameters may correspond to the end of the developing free jet and start of a developed one. [0108] In various embodiments, the effect of jet velocity may vary based on at least one of: the combination of height-to-diameter ratio, radial location and Reynolds number. In various embodiments, a power law of the local Nusselt number may be defined in an equation as follows:

Nu = kRe^a (Eqn. 1) where k and a are constants that depend on height-to-diameter ratio and radial location. In various embodiments, an empirical correlation of these constants may be obtained by experimental work. Figs. 18A-18B show a graphical correlation of both k and a at various conditions. In various embodiments, the results may be obtained for a turbulent jet with Reynolds number ranging from 5,000 to 124,000. In various embodiments, for Jet Reynolds number of 2,000 to 400,000 and height-to-diameter ratio range of 2 to 12, average Nusselt number in radial locations from 2.5 to 7.9 nozzle diameters are found using the following equation:

(Eqn. 2)

[0109] In various embodiments, the problem of mismatch between jet and ambient temperature or concentration may be solved by introducing a recovery (or an adiabatic wall) temperature. In various embodiments, correlations of local heat transfer coefficients may be developed based on the difference between recovery temperature and the surface temperature. In various embodiments, the recovery temperature may take into account the entrainment resulting from the diffusive exchange between the jet and the ambience.

[0110] In various embodiments, the term two-phase flow may include but not limited to cases where a phase change takes place. In various embodiments, in jet impingement, applications that can be described as two phase flows include: drying, spray/mist cooling, and/or nucleate boiling.

[0111] Drying using jet impingement has been used in food industry, textile industry and other applications. In various embodiments, for moderate evaporation rate cases, a heat and mass transfer analogy may be sufficient to predict mass transfer rates from heat transfer ones. In various embodiments Nusselt number may be predicted from Sherwood number using the following analogy equation:

Nu Sh

Rg 0.42 Rg 0.42 (Eqn. 3)

[0112] In various embodiments, Eqns. 1 and 2 can be used for both Nusselt and Sherwood numbers interchangeably. In various embodiments, this application of Eqns. 1 and 2 may be valid if the heat and mass transfer are decoupled. In various embodiments, such in the case of high evaporation rates or higher density variations, a deviation from the heat and mass transfer analogy may occur.

[0113] In various embodiments, another application where jet impingement is considered is in flow boiling. In various embodiments, because in the developing nucleate boiling regime both convective and nucleation heat transfer rates are high, heat transfer rates may be significant compared to pool boiling cases. In various embodiments, in free jet impingement boiling, a saturated or subcooled liquid jet is impinged on a heated surface in a quiescent gas surrounding, while a submerged jet may be characterized by similar jet and surrounding liquids. In various embodiments, in the case of free jets, the jet parameters, such as jet velocity, diameter and subcooling may not be involved in the fully developed nucleate boiling regime. In various embodiments, the effects of such parameters is clear in the single-phase region, developing nucleate boiling region, onset of nucleate boiling (ONB), and critical heat flux (CHF). In various embodiments, correlations may be developed to estimate the complete boiling curve. In various embodiments, as for submerged jet impingement boiling, the jet parameters may affect the fully developed nucleate boiling region. In various embodiments, the effect of the surrounding subcooling may influence the entire process. In various embodiments, another factor influencing jet impingement boiling may include surface condition. In various embodiments, even though single-phase region is not influence by the surface wettability, it is a highly controlling parameter in all the other regions. In various embodiments, lower surface wettability enhances the bubble generation and departure. In various embodiments, lower surface wettability enhances the mixing mechanism that is essential in nucleate boiling. In various embodiments, some experiments were performed on highly conductive heaters, hence constant surface superheat. In addition, heater dimension may be similar to the jet dimension which in turn limits the cases to the stagnation region. In various embodiments, the heater area is greater than the jet diameter. In various embodiments, for lower conductive heaters or heaters with large areas, the heater are appropriately described in constant heat flux terms. In various embodiments, this may result in a variation of the surface temperature in the radial direction with the lowest temperature being at center of the stagnation area. In various embodiments, single phase region, developing and developed nucleate boiling could be observed simultaneously from the center of the stagnation region and radially outward, in the same order. In various embodiments, the ONB may be formed in a shape of a ring with a stable reproducible size.

[0114] In various embodiments, researchers have considered numerically studying such behavior. In various embodiments, numerous numerical models may include Eulerian mixture models, Eulerian mechanistic model, and/or single-phase model. In various embodiments, the Eulerian mixture models are based on numerically solving for the state variables in the vapor and liquid domains separately. In various embodiments, empirical relations were used for the evaporation rates for inter-phase mass transfer. In various embodiments, forces on bubbles were performed by simple drag force and surface tension balance. In various embodiments, the interface heat transfer was set to be infinite therefore constant temperature can be employed at the interface of the two phases. In various embodiments, the Eulerian mechanistic model is similar to that developed for pool boiling. The general form of the mechanistic model is

[0115] In various embodiments, in the fully developed nucleate boiling region, the evaporative heat flux may be neglected. In various embodiments, by knowing the bubble departure diameter ( D ), departure frequency ( ), bubble covered area ratio (A_/,), and nucleation site density (N), the wall heat flux may be predicted. In various embodiments, because of the negligible effect of evaporative heat flux, and the fact that heat transfer is enhanced by the mixing phenomenon caused by bubble departure, the single-phase model may be more appealing. In various embodiments, a modification made to normally solving for state variables is an additional artificial turbulent diffusivity. In various embodiments, jet impingement may be used in spray or mist cooling technology. In various embodiments, in the case of spray cooling, micro-jets may be sprayed directly on a hot surfaces. In various embodiments, high temperature steam may be expanded abruptly and therefore, condensate droplets are generated and impinged on a hot surface in mist cooling. In various embodiments, evaporative cooling is the main mechanism by which heat is removed from the surface.

[0116] In various embodiments, a jet impingement technique may be used to study droplet growth mechanisms on hydrophobic surfaces. In various embodiments, oblique jets may be utilized where the jet is not normal to the surface to reduce its shearing effect. In various embodiments, oblique jets may be used for better visualization of the transient droplet growth. In various embodiments, a solution to a sudden NCG leakage in a pure vapor condensation heat exchanger may include generating a jet of pure steam and impinging the breath on the diffusion layer. In various embodiments, improvements may be obtained of around two fold compared to the absence of the jet. [0117] Fig. 19 illustrates a control volume (CV) over which thermodynamic analysis may be performed. In various embodiments, humid air enters the CV at a temperature equivalent to room temperature (Tl) and relative humidity of (RH1). In various embodiments, the contained water vapor is dehumidified by impingement on a surface at a temperature lower than the dew point. The dehumidified air then leaves the CV with a minute change to its overall mass flow rate. In various embodiments, the condensation process is represented as an energy stored in the system. Because the condensation latent heat (/ ₅~2.447 x 10⁶) is two orders of magnitude higher than sensible heat (c_p T_max~ 1.8 x 10⁴), the latter can be neglected. Therefore, utilizing the condensation rate in the previous section, the heat transfer rate is found as: q" = ™c' h_f3 (Eqn. 4)

Consequently, the overall heat transfer coefficient is obtained as: u = Q _J! kNu

(Eqn. 5)

(^{' T}j^~TRTD ) ^D

[0118] In traditional heat/mass exchangers, the compactness of such machines may be calculated as the ratio between their surface area to their occupied volume which is sometimes referred to as area density. However, this definition lacks an important parameter which is the heat/mass transfer coefficient. Because heat/mass exchanger compactness is a design coefficient to be maximized, the following definition may be more suitable instead of area density: r rh A h_mA

^L,m (Eqn. 6) Dw¥- V q" A _ UA

CH (Eqn. 7) ATV — V

In this case, the heat/mass exchanger system can be estimated to be that of a cylindrical shape. In various embodiments, the base area may be used for normalizing the condensation rate. The height of the cylinder is the nozzle-to-surface spacing (H). In various embodiments, the area density may reduce to (1/H).

[0119] In various embodiments, the EB apparatus may be used as a baseline platform technology in sentinel systems for environmental sampling. In various embodiments, an advantage of the apparatus disclosed herein is that it can be easily modified to sample and condense water vapor from the air as a means of continuously or intermittently monitoring the viral load in critical environments such as hospitals, schools, airports and military bases. In various embodiments, the collection device may port environmental samples to a continuous detection system such as a mass spectrometer. In various embodiments, the compact collector apparatus with a surface modified collection plate and fluidic capture may be piped into existing air quality monitoring systems. In various embodiments, small handheld/portable CDC-based systems may be placed in areas of high concern (ICUs) for local sampling as needed.

[0120] Fig. 20 illustrates an exemplary system 1600 for detecting viral presence. As shown in Fig. 20, the system 1600 may include a dehumidification device and/or an air purifier device. In various embodiments, the system 1600 may force ambient air into a sample collection chamber via one or more fans. In various embodiments, the system 1600 may passively receive ambient air.

[0121] In various embodiments, the system 1600 may include an integral UV light. In various embodiments, the system 1600 may include a camera system ( e.g ., digital camera) for detecting fluorescence (and/or lack thereof). In various embodiments, the system may include an enzyme-linked immunosorbent assay (ELISA). In various embodiments, the system 1600 may include a rapid antigen test (e.g., an immunochromatographi c/lateral flow assay). In various embodiments, as virus in ambient air lands on a chip, the virus may bind to an antibody coating on the sample collection surface. In various embodiments, the bound virus may inactivate a fluorophore (such that the camera would be looking for an absence of fluorescence). In various embodiments, after a predetermined limit of virus is determined, the system 1600 may provide an indication to a user (e.g, throw an alarm, illuminate a light, etc.). In various embodiments, a user may insert/remove a consumable lab-on-a-chip into/from the system 1600.

[0122] In various embodiments, a user may remove the collection material for testing (e.g, at a PCR machine or enzyme-linked immunosorbent assay). In various embodiments, a user may remove the collection material for testing via an antigen test (e.g, lateral flow) used to test for virus presence. In various embodiments, after the collection material has been removed, new collection material may be inserted to replace the spent collection material.

[0123] From the description herein, it will be appreciated that that the present disclosure encompasses multiple embodiments which include, but are not limited to, the following: [0124] An exhaled breath sample collection apparatus, comprising: (a) a sample collection substrate having a surface treated with a hydrophobic material; (b) a collection ring positioned on the surface of the sample collection substrate, said collection ring comprising a hydrophilic material or a functionalized hydrogel; (c) a housing configured to enclose the sample collection substrate and thereby form a sample collection chamber; (d) mouthpiece comprising a tube having a proximal opening and a distal opening, the distal opening in communication with the sample collection chamber, the proximal opening configured to receive exhaled breath from a patient and direct the patient's exhaled breath toward the sample collection substrate such that it impinges on the surface of the sample collection substrate; (e) a one-way valve positioned in the tube to prevent backflow toward the patient; (f) a removable cap over the mouthpiece; (g) one or more one-way valves in the housing that allow exhaled breath to exit the housing after impinging on the surface of the sample collection substrate; (h) a port in the housing with a removable cover that allows insertion of a swab into the sample collection chamber or connection to a processing for in-situ detection of viral load; and (i) a Peltier cooling device positioned beneath the sample collection substrate, said cooling device configured to provide sufficient cooling to condense moisture in the patient's exhaled breath that impinges on the collection surface. [0125] The apparatus of any preceding embodiment, wherein the housing has transparent sidewalls.

[0126] The apparatus of any preceding embodiment, wherein the apparatus is disposable. [0127] The apparatus of any preceding embodiment, wherein the apparatus is a component of an in-situ collection and diagnostic system.

[0128] The descriptions of the various embodiments of the present invention have been presented for purposes of illustration, but are not intended to be exhaustive or limited to the embodiments disclosed. Many modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The terminology used herein was chosen to best explain the principles of the embodiments, the practical application or technical improvement over technologies found in the marketplace, or to enable others of ordinary skill in the art to understand the embodiments disclosed herein.

Claims

What is claimed is:

1. An apparatus comprising: a housing comprising a base and a cover, the base having a sample collection surface, the cover having a port and substantially enclosing the sample collection surface thereby defining a sample collection chamber; a tube extending from a proximal end to a distal end, the proximal end having a proximal opening and the distal end having a distal opening, the tube extending through the port such that the distal opening is in fluid communication with the sample collection chamber, wherein the proximal opening is configured to receive a gaseous sample containing moisture and direct the gaseous sample to the sample collection surface; a cooling device configured to contact the base and cool the sample collection surface to thereby condense at least a portion of the moisture on the sample collection surface; and a sample collection material disposed on the sample collection surface, the sample collection material comprising a hydrogel configured to absorb the condensed moisture.

2. The apparatus of claim 1, further comprising a one-way valve disposed within the tube.

3. The apparatus of claim 1, wherein the hydrogel is functionalized with one or more molecule capable of binding to one or more pathogen.

4. The apparatus of claim 3, wherein the one or more molecule is a receptor configured to bind to the one or more pathogen.

5. The apparatus of claim 3, wherein the one or more molecule is an aptamer configured to bind to the one or more pathogen.

6. The apparatus of claim 1, wherein the hydrogel comprises a covalent conjugation of gelatin and methacrylic anhydride to thereby form a modified gelatin.

7. The apparatus of claim 6, wherein the modified gelatin is functionalized with an enzyme.

8 The apparatus of claim 7, wherein the enzyme comprises ACE2.

9. The apparatus of claim 7 or 8, wherein the enzyme is attached to the modified gelatin by one or more linking molecules.

10. The apparatus of claim 9, wherein the one or more linking molecules comprises an amine.

11. The apparatus of claim 9, wherein the one or more linking molecules comprises an imine.

12. The apparatus of claim 9, wherein the one or more linking molecules comprises glutaric dialdehyde.

13. The apparatus of any one of claim 7 to 12, wherein the sample collection material comprises a first layer and a second layer disposed on top of the first layer, the first layer including the modified gelatin having the enzyme, and the second layer including the modified gelatin without the enzyme.

14. The apparatus of any one of claim 7 to 13, wherein the modified gelatin comprises a photoinitiator configured to photopolymerize the modified gelatin when exposed to visible light.

15. The apparatus of any one of claims 1 to 14, wherein the cover is transparent.

16. The apparatus of any one of claims 1 to 14, wherein the cover comprises a cylindrical shape.

17. The apparatus of any one of claims 1 to 14, wherein the cover comprises an Erlenmeyer flask shape.

18. The apparatus of claim 16 or 17, wherein the cover comprises one or more vent disposed about a circumference of the cover.

19. The apparatus of claim 18, wherein the one or more vent comprises a one-way valve.

20. The apparatus of any one of claims 1 to 19, wherein the sample collection material comprises a ring.

21. The apparatus of claim 20, wherein the ring is positioned on the sample collection surface such that the tube is directed at a center of the ring.

22. The apparatus of claim 20, wherein the sample collection surface comprises a hydrophobic treatment.

23. The apparatus of claim 22, wherein the hydrophobic treatment is applied to an area of the sample collection surface within the ring.

24. The apparatus of claim 22, wherein the hydrophobic treatment comprises a hydrophobic silicon polymer.

25. The apparatus of any one of claims 1 to 24, wherein the cooling device comprises a thermoelectric device.

26. The apparatus of claim 25, wherein the cooling device is thermally coupled to the sample collection surface.

27. The apparatus of any one of claims 1 to 26, wherein a height between the distal end of the tube and the sample collection surface is about 1.5mm to about 52.5mm.

28. The apparatus of any one of claims 1 to 27, wherein a diameter of the tube is about 2mm to about 3mm.

29. The apparatus of any one of claims 1 to 28, wherein a diameter of the cover is at least 10 times a diameter of the tube.

30. The apparatus of any one of claims 1 to 29, wherein a distance between the distal end of the tube and the sample collection surface is about 3 to 5 times a diameter of the tube.

31. The apparatus of any one of claims 1 to 30, wherein the cooling device is configured to cool the sample collection surface to between about 5°C and about 22°C.

32. The apparatus of any one of claims 1 to 31, wherein the apparatus is single-use.

33. An exhaled breath sample collection device comprising: a base having a sample collection surface, the sample collection surface configured to receive a gaseous sample containing moisture and cool the gaseous sample to thereby condense at least a portion of the moisture on the sample collection surface; and a sample collection material disposed on the sample collection surface, the sample collection material comprising a hydrogel configured to absorb the condensed moisture.

34. The device of claim 33, wherein the sample collection surface comprises a circular area.

35. The device of claim 33 or 34, wherein the sample collection material comprises a ring.

36. The device of claim 35, wherein a center of the ring is at a center of the circular area.

37. The device of claim 33, wherein the hydrogel is functionalized with one or more molecule capable of binding to one or more pathogens.

38. The device of claim 37, wherein the one or more molecule is a receptor configured to bind to the one or more pathogen.

39. The device of claim 37, wherein the one or more molecule is an aptamer configured to bind to the one or more pathogen.

40. The device of any one of claims 33 to 39, wherein the hydrogel comprises a covalent conjugation of gelatin and methacrylic anhydride to thereby form a modified gelatin.

41. The device of claim 40, wherein the modified gelatin is functionalized with an enzyme.

42. The device of claim 41, wherein the enzyme comprises ACE2.

43. The device of claim 41 or 42, wherein the enzyme is attached to the modified gelatin by one or more linking molecules.

44. The device of claim 43, wherein the one or more linking molecules comprises an amine.

45. The device of claim 43, wherein the one or more linking molecules comprises an imine.

46. The device of claim 43, wherein the one or more linking molecules comprises glutaric dialdehyde.

47. The device of any one of claim 41 to 46, wherein the sample collection material comprises a first layer and a second layer disposed on top of the first layer, the first layer including the modified gelatin having the enzyme, and the second layer including the modified gelatin without the enzyme.

48. The device of claim 33, wherein the base comprises a material capable of remaining cool for about 2 minutes to thereby condense moisture contained with a sample of human breath directed at the sample collection surface.

49. The device of claim 33, wherein the base comprises a material having a ratio of thermal conductivity to length greater than 300 W/m^A2K.

50. The device of claim 33, wherein the base comprises a material having a specific heat greater than 1.5 kJ/kg/K.

51. The device of claim 33, wherein the base comprises a material having an effusivity greater than 25 J/m^A2/K/s^Al/2.

52. The device of claim 33, wherein the device is single-use.

53. The device of any one of claim 41 to 52, wherein the modified gelatin comprises a photoinitiator configured to photopolymerize the modified gelatin when exposed to visible light.

54. A method for forming an exhaled breath sample collection device, the method comprising: applying a mask to a first portion of a substrate such that a second portion of the substrate does not include the mask; applying a hydrogel solution to the second portion, wherein the hydrogel solution comprises a hydrogel and a photoinitiator; photocrosslinking the hydrogel solution; and removing the mask.

55. The method of claim 54, wherein the second portion forms a ring on the substrate

56. The method of claim 54, wherein applying the hydrogel solution comprises spin coating.

57. The method of claim 54, wherein the hydrogel comprises a covalent conjugation of gelatin and methacrylic anhydride to thereby form a modified gelatin.

58. The method of claim 57, wherein the modified gelatin is functionalized with an enzyme.

59. The method of claim 58, wherein the enzyme comprises ACE2.

60. The method of claim 58 or 59, wherein the enzyme is attached to the modified gelatin by one or more linking molecules.

61. The method of claim 60, wherein the one or more linking molecules comprises an amine.

62. The method of claim 60, wherein the one or more linking molecules comprises an imine.

63. The method of claim 60, wherein the one or more linking molecules comprises glutaric dialdehyde.

64. The method of any one of claim 57 to 63, wherein applying the hydrogel solution comprises applying a first layer including the modified gelatin having the enzyme; and wherein photocrosslinking the hydrogel solution comprises photocrosslinking the first layer.

65. The method of claim 64, further comprising applying a second layer disposed on top of the first layer, the second layer including the modified gelatin without the enzyme; and photocrosslinking the second layer.

66. A method for detecting a pathogen contained within breath of a human, the method comprising: receiving a gaseous sample of human breath containing moisture at a sample collection surface on a base; cooling the sample collection surface to a temperature lower than the dew point of the moisture in the gaseous sample thereby condensing the moisture on the sample collection surface; and collecting at least a portion of the condensed moisture at a sample collection material disposed on the sample collection surface, the sample collection material comprising a hydrogel.

67. The method of claim 66, wherein receiving the gaseous sample comprises receiving the gaseous sample at a flow rate of about 1 liter per minute (LPM) to about 3 LPM.

68. The method of claim 66, wherein the sample collection material comprises a ring.

69. The method of claim 66, wherein the sample collection surface comprises a hydrophobic treatment applied to an area of the sample collection surface within the ring.

70. The method of claim 69, wherein the hydrophobic treatment comprises a hydrophobic silicon polymer.

71. The method of claim 66, wherein receiving the gaseous sample comprises providing a tube configured to receive the human breath.

72. The method of claim 66, wherein the cooling device comprises a thermoelectric device.

73. The method of claim 72, wherein the cooling device is thermally coupled to the sample collection surface.

74. The method of claim 66, further comprising removing the base from the cooling device prior to receiving the gaseous sample.

75. The method of claim 66, wherein the base comprises a material capable of remaining cool for about 2 minutes to thereby condense moisture contained with a sample of human breath directed at the sample collection surface.

76. The method of claim 66, wherein the base comprises a material having a ratio of thermal conductivity to length greater than 300 W/m^A2K.

77. The method of claim 66, wherein the base comprises a material having a specific heat greater than 1.5 kJ/kg/K.

78. The method of claim 66, wherein the base comprises a material having an effusivity greater than 25 J/m^A2/K/s^Al/2.

79. The method of any one of claims 66 to 74, further comprising processing the collected condensed moisture to thereby determine a presence of a pathogen contained within the moisture.

80. The method of claim 79, wherein processing the collected condensed moisture comprises: transferring the sample collection surface to a fluorescence in situ hybridization (FISH) developer; and fixing cells within the collected condensed moisture; applying FISH probes to the collected condensed moisture; and washing the sample collection material.

81. The method of claim 80, wherein fixing the cells comprises applying methanol or ethanol to the sample collection material.

82. The method of claim 80, wherein the FISH probes comprise RNA probes.

83. The method of claim 80, wherein the FISH probes are configured to attach to RNA within the pathogen.

84. The method of claim 83, wherein the pathogen comprises SARS-CoV-2.

85. The method of any one of claims 80 to 84, further comprising imaging the processed condensed moisture with a wide-area fluorescence scanner to thereby detect the presence of the pathogen.

86. A kit comprising: the device of any one of claims 33-53; and a swab having at least one frangible region along a swab shaft.

87. An apparatus comprising: a housing comprising a base and a cover, the base having a sample collection surface, the cover having a port and substantially enclosing the sample collection surface thereby defining a sample collection chamber; a tube extending from a proximal end to a distal end, the proximal end having a proximal opening and the distal end having a distal opening, the tube extending through the port such that the distal opening is in fluid communication with the sample collection chamber, wherein the proximal opening is configured to receive a gaseous sample containing moisture and direct the gaseous sample to the sample collection surface; a cooling device configured to contact the base and cool the sample collection surface to thereby condense at least a portion of the moisture on the sample collection surface; and one or more microfluidic channels disposed within the base, the one or more microfluidic channels being configured to collect the condensed moisture.

88. The apparatus of claim 87, wherein the one or more microfluidic channels comprises a ring.

89. The apparatus of claim 88, wherein the one or more microfluidic channels comprises an outflow channel extending radially outward from the ring.

90. The apparatus of claim 87, wherein the one or more microfluidic channels comprises a central area.

91. The apparatus of claim 90, wherein the central area is substantially circular.

92. The apparatus of claim 90, wherein the one or more microfluidic channels further comprises an outer ring.

93. The apparatus of claim 92, further comprising two or more spoke channels connecting the central area to the outer ring.

94. The apparatus of claim 87, wherein the base comprises a first layer and a second layer above the first layer.

95. The apparatus of claim 94, wherein the first layer comprises a hydrophilic-coated substrate.

96. The apparatus of claim 94, wherein the first layer comprises a glass substrate.

97. The apparatus of claim 94, wherein the first layer comprises HDPE.

98. The apparatus of claim 94, wherein the second layer comprises a treated coating.

99. The apparatus of claim 98, wherein the treated coating comprises a hydrophobic coating.

100. The apparatus of claim 87, wherein the one or more microfluidic channels comprises a width of about 1/16 inch to about 3/16 inch.

101. A method for detecting a virus contained within breath of a human, the method comprising: receiving a gaseous sample of human breath containing moisture at a sample collection surface; cooling the sample collection surface to a temperature lower than the dew point of the moisture in the gaseous sample thereby condensing the moisture on the sample collection surface; and collecting at least a portion of the condensed moisture within a microfluidic channel formed in the sample collection surface.

102. The method of claim 101, further comprising adding a transport medium into the collected condensed moisture.

103. The method of claim 101 or 102, further comprising transferring the collected condensed moisture into a container.

104. The method of claim 101, further comprising processing the condensed moisture, wherein processing the collected condensed moisture comprises: transferring the sample collection surface to a fluorescence in situ hybridization (FISH) developer; and fixing cells within the collected condensed moisture; applying FISH probes to the collected condensed moisture; and washing the sample collection material.

105. The apparatus of any one of claims 1 to 32 or 87 to 100, wherein a thickness of the base is selected such that the base can cool from about 20°C to 5°C in less than 10 seconds after being placed on the cooling device.

106. The apparatus of any one of claims 1 to 32 or 87 to 100, wherein the base comprises a material capable of remaining cool for about 2 minutes to thereby condense moisture contained with a sample of human breath directed at the sample collection surface.

107. The apparatus of any one of claims 1 to 32 or 87 to 100, wherein the base comprises a material having a ratio of thermal conductivity to length greater than 300 W/m^A2K.

108. The apparatus of any one of claims 1 to 32 or 87 to 100, wherein the base comprises a material having a specific heat greater than 1.5 kJ/kg/K.

109. The apparatus of any one of claims 1 to 32 or 87 to 100, wherein the base comprises a material having an effusivity greater than 25 J/m^A2/K/s^Al/2.