WO2023014842A1

WO2023014842A1 - Syringe and components thereof

Info

Publication number: WO2023014842A1
Application number: PCT/US2022/039345
Authority: WO
Inventors: Dimitri A. Bikos; David A. Weitz; Michael R. Pavia; Thomas B. LEFEVRE; Isaak J. THORNTON; James N. Wilking
Original assignee: President And Fellows Of Harvard College; Montana State University
Priority date: 2021-08-04
Filing date: 2022-08-03
Publication date: 2023-02-09

Abstract

An integrated device for controlled sampling and detection can include a vial containing a reagent. The device can include a tube. The tube can include a plurality of prongs configured to collect a predetermined amount of sample and to pierce a seal of vial. The device can include a blister pack containing a liquid. The device can include a plunger configured to cause the blister pack to release the liquid toward the sample and to direct the sample into the vial.

Description

SYRINGE AND COMPONENTS THEREOF

CROSS-REFERENCE TO RELATED PATENT APPLICATION

[0001] This application claims the benefit and priority of U.S. Provisional Patent Application No. 63/229,472, filed on August 4, 2021, the entirety of which is incorporated by reference herein.

TECHNICAL FIELD

[0002] The present application relates generally to rapid detection of pathogens.

BACKGROUND

[0003] Infectious diseases can be caused by pathogens such as bacteria, fungi, protozoa, worms, viruses, and prions.

SUMMARY

[0004] The systems and methods of the present disclosure are directed to a device that can detect low levels of a pathogen rapidly, inexpensively, and without the need for trained personnel, personal protective equipment, and/or highly specialized equipment. The device and associated assay can provide a widely deployable, rapid screening test. The device, in some embodiments, can detect SARS-CoV-2 (or other virus) and provide widespread, rapid, and inexpensive testing with the sensitivity of current “gold-standard” tests using quantitative polymerase chain reaction (qPCR). The device can be used for any other nucleic acid test, including for other viruses, bacteria, fungi, or other nucleic acid targets. For example, the device can be used for flu, Ebola, any SARS species, MERS, or any other viral or bacterial species. The device can be used to screen individuals entering a school, a university, a sports venue, a theater, a concert, an office building, an airport, or a restaurant. The results of the test can be combined with a smartphone-based application.

[0005] Human microbial pathogens including viruses, bacteria, and fungi can pose a profound threat to human health, as evidenced by the SARS-CoV-2 global pandemic. To limit the spread of infectious disease, frequent, widespread diagnostic testing can be needed. Specifically, there can be a need for low-cost, accurate molecular assays (e.g., point-of-care assays, clinical lab assays). Isothermal amplification methods can offer a potential solution, and molecular-based point-of-care (POC) tests can offer potential for rapid, accurate POC testing, but to keep reagent costs low, methods for aliquoting exact volumes of specimen sample onto the assay device can be needed. The systems and methods of the present disclosure are directed to low-cost, hand-held, manually operated device for rapid pathogen detection. The device can be used, for example, in a POC setting or in a clinical lab setting. The device can include a two-pronged fork geometry which can enable transfer of small volumes of saliva from flocked collection swabs and can include reagents needed for isothermal nucleic acid amplification and fluorescence detection. The design and operation of the device can be describe and use of the device for isothermal nucleic acid amplification and fluorescence detection using loop-mediated isothermal amplification of the SARS-CoV-2 virus in saliva can be demonstrated.

[0006] At least one aspect of the present disclosure is directed to an integrated device for controlled sampling and detection. The integrated device can include a vial containing a reagent. The integrated device can include a tube. The tube can include a plurality of prongs. The plurality of prongs can collect a predetermined amount of sample. The plurality of prongs can pierce a seal of the vial. The integrated device can include a blister pack containing a liquid. The integrated device can include a plunger configured to cause the blister pack to release the liquid toward the sample to direct the sample into the vial.

[0007] Another aspect of the present disclosure is directed to an integrated device for controlled sampling and detection. The integrated device can include a vial containing a reagent. The integrated device can include a tube. The tube can include an aperture configured to collect a predetermined amount of sample. The tube can include a portion configured to pierce a seal of the vial. The integrated device can include a blister pack containing a liquid. The integrated device can include a plunger configured to cause the blister pack to release the liquid toward the sample to direct the sample into the vial.

[0008] Another aspect of the present disclosure is directed to a method of collecting and providing a sample. The method can include providing a vial containing a reagent, a tube that includes a plurality of prongs, a blister pack containing a liquid, a plunger, and/or a flocked swab that includes the sample. The method can include collecting a predetermined amount of the sample into the tube by drawing the flocked swab between the plurality of prongs. The method can include piercing the vial with the plurality of prongs. The method can include pressing the plunger into the tube to cause the blister pack to release the liquid toward the sample to direct the sample into the vial.

[0009] Those skilled in the art will appreciate that the summary is illustrative only and is not intended to be in any way limiting. Other aspects, inventive features, and advantages of the devices and/or processes described herein, as defined solely by the claims, will become apparent in the detailed description set forth herein and taken in conjunction with the accompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

[0010] The details of one or more implementations of the subject matter described in this specification are set forth in the accompanying drawings and the description below.

Other features, aspects, and advantages of the subject matter will become apparent from the description, the drawings, and the claims.

[0011] FIG. 1 illustrates a perspective view of an integrated device for controlled sampling and detection, according to an embodiment.

[0012] FIG. 2A illustrates a cross-sectional view of the integrated device for controlled sampling and detection shown in FIG. 1 taken along plane A-A, according to an embodiment.

[0013] FIG. 2B illustrates a perspective view of a kit, according to an embodiment.

[0014] FIG. 3 illustrates an exploded view of the integrated device for controlled sampling and detection, according to an embodiment.

[0015] FIG. 4 illustrates a perspective view of a tube, according to an embodiment.

[0016] FIG. 5 illustrates a perspective view of the tube, according to an embodiment.

[0017] FIG. 6 illustrates a perspective view of the tube, according to an embodiment.

[0018] FIG. 7 illustrates a perspective view of the tube, according to an embodiment.

[0019] FIG. 8 illustrates a cross-sectional view of the tube shown in FIG. 7 taken along plane B-B, according to an embodiment.

[0020] FIG. 9 illustrates a perspective view of a plunger, according to an embodiment.

[0021] FIG. 10 illustrates a method of collecting and providing a sample, according to an embodiment.

[0022] FIG. 11 A illustrates a side view of prongs and a front view of the prongs, according to an embodiment. [0023] FIG. 1 IB illustrates a plot of volume of saliva transferred between the prongs vs. saliva volume in a swab, according to an embodiment.

[0024] FIG. 11C illustrates a histogram of saliva volume transferred across multiple individuals, according to an embodiment.

[0025] FIG. 1 ID illustrates a time lapse of a process for transferring saliva, according to an embodiment.

[0026] FIG. 12A illustrates a schematic of depressions of a plunger, according to an embodiment.

[0027] FIG. 12B illustrates a diagram of depression of the plunger, blister compression, and expulsion of a liquid, according to an embodiment.

[0028] FIG. 12C illustrates a plot of force vs. displacement, according to an embodiment.

[0029] FIG. 12D illustrates a plot of force vs. blister compression fraction, according to an embodiment.

[0030] Like reference numbers and designations in the various drawings indicate like elements.

DETAILED DESCRIPTION

[0031] Following below are more detailed descriptions of various concepts related to, and implementations of, methods, apparatuses, and systems for controlled sampling and detection of pathogens. The various concepts introduced above and discussed in greater detail below may be implemented in any of a number of ways, as the described concepts are not limited to any particular manner of implementation. Examples of specific implementations and applications are provided primarily for illustrative purposes.

[0032] Embodiments herein relate generally to integrated devices for controlled sampling and detection, and methods for collecting and providing a sample. Particularly, various embodiments described herein provide a two-part device. The device can include (1) an upper portion containing both the aqueous solvent and a region into which a sample is loaded with a collection swab (e.g., flocked swab), and (2) a lower portion containing amplification reagents. The sample can be collected by drawing a sample collection swab through two narrow forks (e.g., prongs) in the upper portion of the device. The upper portion of the device and the lower portion of the device can then be assembled. A plunger in the upper portion can be depressed, thus driving the flow of aqueous solvent and sample into a chamber in the lower portion of the device. Detection and determination of a viral load can be achieved through an amplification step followed by a detection step. The amplification can be performed through loop mediated isothermal amplification (LAMP), using fluorescent detection. The sample and reagents can be heated by placing the entire assembly into a heater at 65°C for 30 minutes, for example. The sample and reagents can undergo an amplification method, for example, a LAMP assay, qPCR, or other PCR method. The fluorescent signal from the amplified template can be detected with a small box that includes a sample holder to hold the device, LEDs to excite the fluorescence, filters to block the excitation light, a camera to detect the fluorescent signal from the sample volume, and/or a heater or heat cycler. The viral load can be determined by measuring the intensity of the fluorescence against a fluorescent reference dye.

[0033] Various embodiments described herein may provide one or more benefits including, for example: (1) providing a widely deployable rapid screening test for the detection of viruses and the determination of viral loads; (2) providing a low-cost kit for the screening test as compared with conventional sample collection and detection kits; (3) providing a device for collecting and introducing a clinical sample to a detection chamber; (4) providing a device for controlled sampling and detection of viruses or pathogens; (5) providing a device with structural components configured to collect a controlled amount of a sample and deliver the sample into a test chamber; (6) providing a device that allows sterile water in a contained environment to be introduced to the test chamber to activate test reagents; and (7) providing a method for collecting and providing a controlled amount of sample to the test chamber.

[0034] Reverse transcription loop-mediated isothermal amplification (RT-LAMP) can include an isothermal, rapid technique for the exponential amplification of RNA that avoids the thermal cycling and high temperatures of reverse transcription polymerase chain reaction (RT-qPCR). RT-LAMP can use a set of four to six unique primers that, in the presence of DNA polymerase, repeatedly produce copies of target cDNA through the formation of loop structures. Replacing PCR with LAMP amplification can represent a step forward for molecular nucleic acid testing. At the clinical laboratory test scale, the RT-qPCR test is considered a complex test where results can only be obtained by trained personnel. Current processing using the CDC approved diagnostic panel for RT-qPCR can take at least 3 hours, and usually much longer, to complete, including sample acquisition, RNA extraction, reagent preparation, and analysis of the viral Nl, N2 and human RNase P genes. In contrast, LAMP can amplify a single copy of RNA to 10⁹ in less than an hour. LAMP amplification can be much faster than PCR, even when there are large amounts of non-target RNA present, and even with “dirty” samples such as blood, serum, feces, and nasal swabs. There is no need for equipment except a water bath or heat block at a temperature of 65°C for LAMP, which allows LAMP to avoid the thermal cycling and high temperatures required by the gold standard RT-qPCR, which takes two hours to run. By replacing PCR with LAMP chemistry, significant gains in turnaround time can be achieved.

[0035] The integrated device can be based upon bulk LAMP amplification. The sample collection geometry can allow for collection of a small quantity of sample, for example, between 2 and 15 pL, which can limit the amount of LAMP reagents needed for each test. This can keep the cost of the reaction low. The systems and methods of the present disclosure can achieve high throughput testing by using elegantly designed, inexpensive, and disposable devices. Under isothermal conditions, the amplification signal arising from low virion concentrations can be rapidly amplified and detected. The device can be engineered to optimize sample collection geometry and device actuation, and provide high signal-to-noise.

[0036] FIG. 1 illustrates a perspective view of an integrated device 100 (e.g., integrated assembly, two-part device, syringe-type point-of-care device, syringe-type viral load detector, device, LAMP-device, syringe device) for controlled sampling and detection. The integrated device 100 can include a vial 105 (e.g., reaction tube, test chamber, microcentrifuge tube, etc.). The vial 105 can be sealed with a thin film (e.g., membrane, multi -laminate film, thin membrane seal, seal, film, foil, etc.). The vial 105 can include an Eppendorf tube (e.g., 0.5 mL Eppendorf tube). The vial 105 can include a sealed plastic vial. The vial 105 can be covered with the thin film to protect the contents of the vial 105 from the outside environment.

[0037] The vial 105 can include (e.g., contain) a reagent 110. The reagent 110 can include a lyophilized LAMP mixture. The reagent 110 can include a lyophilized bead. The reagent 110 can include LAMP reagents in lyophilized form. The reagent 110 can include a lyophilized reagent. The reagent 110 can include a lyophilized LAMP formulation. The reagent 110 can detect a pathogen (e.g., coronavirus, SARS-CoV-2, SARS, MERS, influenza virus, Ebola virus, or any other viral or bacterial species). The reagent 110 can include primers adapted for a particular pathogen. For example, the LAMP reagent can include a primer for amplification of nucleic acids for the detection of SARS-CoV-2 and SARS-CoV-2 variants (e.g., SARS-CoV-2 Alpha variant, SARS-CoV-2 Beta variant, SARS-CoV-2 Gamma variant, SARS-CoV-2 Delta variant, etc.). The reagent 110 can be isolated from the outside environment by the sealed vial. The reagent 110 can be sensitive to (e.g., formulated, and/or its amount and/or concentration determined according to) the (expected) amount of sample collected.

[0038] The integrated device 100 can include a tube 115 (e.g., syringe tube). The tube 115 can include a cylindrical tube. The tube 115 can include a handle 130. The handle 130 can be gripped. The tube 115 can be inserted into the vial 105. The tube 115 can couple with the vial 105. For example, the tube 115 can couple with the vial 105 via an interference fit (e.g., press fit, friction fit, etc.). One end of the tube 115 can couple with the vial 105. The tube 115 can have a cylindrical shape. The tube 115 can have a tapered shape. The tube 115 can be hollow.

[0039] The tube 115 can be manufactured via injection molding or other fabrication methods. The tube 115 can made of an injection moldable material. The tube 115 can be made of a mechanically stable material, a chemically resistant material, and/or an optically transparent material. For example, the tube 115 be made of a cyclic olefin copolymer. The tube 115 can be made of a biomedical grade polypropylene. The tube 115 can be made of plastic. The tube 115 can be made of a rigid material. The tube 115 can be made of a material that has an air-water contact angle of approximately 90°. The tube 115 can be made of a material that is slightly non-wetting. The tube 115 can be made of a material that is slightly hydrophobic or slightly hydrophilic. The tube 115 can be made of a material that is used for point-of-care diagnostic devices. The tube 115 can be made of a material that is in a clinical lab setting. The tube 115 can be sterilized.

[0040] The tube 115 can include a plurality of prongs 120 (e.g., forks, tines, spikes, puncture points, forked prongs, puncture tips, protrusion, etc.). A swab (e.g., collection swab, flocked swab, flocked specimen swab, standard sampling swab, etc.) can be drawn through/b etween the plurality of prongs 120. The plurality of prongs 120 can be made of a material that is rigid enough to resist deformation as the swab is drawn through the plurality of prongs 120. The plurality of prongs 120 can be made of a material that is rigid enough to compress (e.g., allow for compression of) a flocked end of the swab, e.g., to release a specific amount (e.g., within an expected range, predetermined amount) of sample from the swab to the prongs/tube. The plurality of prongs 120 can be shaped such that the plurality of prongs 120 can puncture the vial 105 or the thin film of the vial 105. The plurality of prongs 120 can be made of a material that is rigid enough to puncture the vial 105 or the thin film of the vial 105. The plurality of prongs 120 can be positioned at one end of the tube 115. The plurality of prongs 120 can receive (e.g., collect) a predetermined (e.g., clinical) amount of the sample, e.g., from the drawn swab. For example, the predetermined amount of the sample can include a volume in a range of about 2 pL to 15 pL (e.g., 2 pL, 3 pL, 4 pL, 5 pL, 6 pL, 7 pL, 8 pL, 9 pL, 10 pL, 11 pL, 12 pL, 13 pL, 14 pL, or 15 pL, inclusive). The plurality of prongs 120 can mete out a precise amount of the sample. A small, precise amount of the sample can reduce (and/or deterministically set or predefine) the amount of reagent 110 for the amplification reaction to take place.

[0041] The plurality of prongs 120 can be manufactured via injection molding or other fabrication methods. The plurality of prongs 120 can made of an injection moldable material. The plurality of prongs 120 can be made of a mechanically stable material, a chemically resistant material, and/or an optically transparent material. For example, the plurality of prongs 120 can be made of a cyclic olefin copolymer. The plurality of prongs 120 can be made of a biomedical grade polypropylene. The plurality of prongs 120 can be made of plastic. The plurality of prongs 120 can be made of a rigid material. The plurality of prongs 120 can be made of a material that has an air- water contact angle of approximately 90°. The plurality of prongs 120 can be made of a material that is slightly non-wetting. The plurality of prongs 120 can be made of a material that is slightly hydrophobic or slightly hydrophilic. The plurality of prongs 120 can be made of the same or different material than the tube 115. The plurality of prongs 120 can be made of a material that is used for point-of- care diagnostic devices. The plurality of prongs 120 can be made of a material that is used in a clinical lab setting. The plurality of prongs 120 can be sterilized.

[0042] The plurality of prongs 120 can couple with the vial 105. For example, the plurality of prongs 120 can couple with the vial 105 via an interference fit (e.g., press fit, friction fit, etc.). The plurality of prongs 120 can couple with the vial 105 to produce a joint (e.g., connection, seal, etc.) which is held together by friction after the plurality of prongs 120 and the vial 105 are pushed together. The vial 105 can be pushed onto the plurality of prongs 120. The plurality of prongs 120 can pierce the thin film of the vial 105. The plurality of prongs 120 can pierce a seal of the vial 105. The plurality of prongs 120 can be pushed into the vial 105. The process of creating an interference fit between the plurality of prongs 120 and the vial 105 can seal and/or reseal the vial 105. Sealing or resealing the vial 105 can isolate the sample (e.g., saliva sample) contained within the vial 105 from the outside environment and possible contamination.

[0043] The integrated device 100 can include a plunger 125 (e.g., blister actuator). The plunger 125 can couple with the tube 115. For example, the plunger 125 can be inserted into the tube 115. The plunger 125 can have a smaller diameter than the tube 115. The plunger 125 can be pushed into the tube 115. The integrated device 100 can have an ergonomic handling length. For example, the handle 130 can be gripped with one or more fingers (e.g., four fingers) and the plunger 125 can be pressed with a thumb. The integrated device 100 can be pre-assembled. For example, the plunger 125 can be partially inserted into the tube 115. The plunger 125 can be inserted into the tube 115 up to a snap lock or other extent. The plunger 125 can have a cylindrical shape. The plunger 125 can have a tapered shape.

[0044] The plunger 125 can be manufactured via injection molding or other fabrication methods. The plunger 125 can made of an injection moldable material. The plunger 125 can be made of a mechanically stable material, a chemically resistant material, and/or an optically transparent material. For example, the plunger 125 be made of a cyclic olefin copolymer. The plunger 125 can be made of a biomedical grade polypropylene (e.g., biomedical grade polypropylene). The plunger 125 can be made of plastic. The plunger 125 can be made of a rigid material. The plunger 125 can be made of a material that has an airwater contact angle of approximately 90°. The plunger 125 can be made of a material that is slightly non-wetting. The plunger 125 can be made of a material that is slightly hydrophobic or slightly hydrophilic. The plunger 125 can be made of the same or different material than the tube 115. The plunger 125 can be made of the same or different material than the plurality of prongs 120. The plunger 125 can be made of a material that is used for point-of-care diagnostic devices. The plunger 125 can be made of a material that is used in a clinical lab setting. The plunger 125 can be sterilized. The plunger 125 can be made of a material that is rigid enough such that it can be driven through the tube 115. The tube 115 can be made of a material that is rigid enough to allow the plunger 125 to be driven through the tube 115.

[0045] FIG. 2 A illustrates a cross-sectional view of an integrated device 100 for controlled sampling and detection (e.g., shown in FIG. 1 taken along plane A-A). The integrated device 100 can include the vial 105, the reagent 110, the tube 115, the plurality of prongs 120, and the plunger 125. The tube 115 can have an inner tube diameter 215. The plunger 125 can have a plunger diameter 220. The plunger diameter 220 can be less than (or greater than, or equal to) the inner tube diameter 215.

[0046] The integrated device 100 can include a blister pack 205 (e.g., sealed blister pack, liquid reservoir, blister, blister dome, etc.). The blister pack 205 can include a liquid- filled blister pack. The blister pack 205 can contain an aqueous solvent (e.g., water, sterile water, etc.). The blister pack 205 can include a liquid. The liquid, the sample, and the reagent 110 can be configured to mix. The blister pack 205 can include a multi-laminate film. For example, a side (e.g., underside) of the blister pack 205 can include a multi-laminate film. Liquid in the blister pack 205 can be driven from the blister pack 205 by gripping the handle 130 with one or more fingers (e.g., four fingers) and pressing the plunger 125 with the thumb. The blister pack 205 can be punctured by the tube 115 as a result of the plunger 125 being pressed into the tube 115. The blister pack 205 can have a volume in a range of about 110 pL to 150 pL (e.g., 110 pL, 120 pL, 130 pL, 140 pL, or 150 pL, inclusive). The blister pack 205 can contain a volume of liquid (e.g., water, sterile water, etc.) in a range of about 30 pL to 50 pL (e.g., 30 pL, 35 pL, 40 pL, 45 pL, or 50 pL, inclusive). ). The blister pack 205 can contain a sufficient amount of liquid to dissolve the reagent 110.

[0047] The integrated device 100 can be pre-assembled. For example, the blister pack 205 can be positioned at an end of the tube 115. The blister pack 205 can be positioned at the end of the tube 115 near the plurality of prongs 120. The blister pack 205 can be inserted into the tube 115. The blister pack 205 can adhere to the cylindrical tube. For example, the blister pack 205 can adhere to a bottom of the tube 115. The tube 115 can include puncture points to puncture the blister pack 205. The tube 115 can include at least one protrusion to puncture the blister pack 205.

[0048] The tube 115 can include an aperture 210 (e.g., hole). The aperture 210 can be at a predetermined, proximate location relative to the plurality of prongs, to draw the sample (e.g., saliva sample) positioned on and/or between the plurality of prongs 120 towards the blister pack 205. The sample can be drawn into the aperture 210 when the swab is drawn through the plurality of prongs 120. The sample can be drawn into the aperture 210 via capillary action. The aperture 210 can collect the predetermined amount of the sample. The aperture 210 can include a cylindrical hole. The aperture 210 can have a circular crosssection. One end of the aperture 210 can be sealed against the atmosphere. The aperture 210 can include an aperture volume. The aperture volume can be large enough relative to the volume of the sample such that the sample is not driven out of the aperture 210. The aperture volume can be large enough to prevent sample expulsion. Air within the aperture 210 can be compressed as the sample is loaded into the aperture 210. The pressure associated with compression of the air can be described by the ideal gas law, PV = nRT. The pressure resisting expulsion of the fluid (e.g., sample) can include the capillary pressure, C_p ~

where <J is the air-saliva interfacial tension and a is the cross-sectional radius of the aperture 210 (e.g., hole).

[0049] The integrated device 100 can include an upper portion (e.g., upper device portion, upper device portion assembly, etc.) and a lower portion (e.g., lower device portion, lower device portion assembly, etc.). The upper portion can include the aqueous solvent and a region into which the sample is loaded with a collection swab. For example, the upper portion can include the blister pack 205, the tube 115, and the plunger 125. The upper portion can be pre-assembled. The upper portion can be prepared for sample collection and actuation. The lower portion can include the reagents for amplification (e.g., amplification reagents, LAMP reagents, etc.). For example, the lower portion can include the vial 105. The lower portion can include a lyophilized LAMP mixture. The lower portion can be sealed with a thin film.

[0050] FIG. 2B illustrates a perspective view of a kit 250 (e.g., test kit, sample collection and detection kits, etc.) can include the integrated device 100. The integrated device 100 can include the vial 105, the reagent 110, the tube 115, the plurality of prongs 120, the plunger 125, the handle 130, and the blister pack 205 (not shown).

[0051] The kit 250 can include a swab 255 (e.g., collection swab, flocked swab, flocked specimen swab, standard sampling swab, sterile swab, sterile flocked swab, applicator swab, etc.). The swab 255 can include a saliva sampling swab to collect saliva or other sample. The saliva/sample can contain surfactants and other surface active molecules that modify the contact angle of the tube 115 and make the saliva/sample wet the tube 115. For example, the swab 255 can be inserted into the lower portion of a patient’s mouth for a time in a range of 5 seconds to 20 seconds (e.g., 5 seconds, 10 seconds, 15 seconds, or 20 seconds, inclusive), or until the swab 255 is soaked with saliva. The swab 255 can include a nasal mucus sampling swab to collect nasal mucus.

[0052] The swab 255 can have an outer swab diameter 260. The outer swab diameter 260 can include the diameter of the sample collection end of the swab 255. The outer swab diameter 260 can be less than the inner tube diameter 215. The outer swab diameter 260 can be less than (or greater than, or equal to) the plunger diameter 220. The swab 255 can have an inner swab diameter 265. The inner swab diameter 265 can include the diameter of the rod end of the swab 255. The inner swab diameter 265 can be less than the outer swab diameter 260. The inner swab diameter 265 can be less than (or greater than, or equal to) the inner tube diameter 215. The inner swab diameter 265 can be less than (or greater than, or equal to) the plunger diameter 220.

[0053] FIG. 3 illustrates an exploded view of the integrated device 100 for controlled sampling and detection. The integrated device 100 can include the vial 105, the reagent 110, the tube 115, the plurality of prongs 120, the plunger 125, and/or the blister pack 205. The vial 105 can include a membrane seal 305 (e.g., thin film, membrane, multi-laminate film, thin membrane seal, seal, film, etc.). The plurality of prongs 120 can pierce (e.g., puncture) the membrane seal 305.

[0054] FIG. 4 illustrates a perspective view of the tube 115. The tube 115 can include the plurality of prongs 120 and the aperture 210. The tube 115 can include a plurality of first facets 405. The plurality of prongs 120 can include the plurality of first facets 405. The plurality of first facets 405 can be angled in such a way that the plurality of prongs 120 are pointed at the ends. The plurality of first facets 405 can be angled in such a way that the plurality of prongs 120 can puncture the membrane seal 305 of the vial 105. The plurality of first facets 405 can include a plurality of bevels. The plurality of prongs 120 can be beveled.

[0055] The tube 115 can include a plurality of second facets 410. The plurality of prongs 120 can include the plurality of second facets 410. The plurality of second facets 410 can be angled in such a way that the sample is drawn into the aperture 210 rather than expelled around the outside of the plurality of prongs 120. For example, the plurality of second facets 410 can be angled in such a way that the sample (or a substantial/specific/configured amount of the sample) is drawn/directed into the aperture 210 rather than expelled around the outside of the plurality of prongs 120 when the swab 255 is drawn through the plurality of prongs 120. The plurality of second facets 410 (e.g., size, direction, shape, and/or surface characteristic of the facets) can guide/direct/funnel the sample towards and into the aperture 210.

[0056] FIG. 5 illustrates a perspective view of the tube 115. The tube 115 can include the plurality of prongs 120 and the aperture 210. Each of the plurality of prongs 120 can be separated by an inner distance 505. The inner distance 505 can be less than the outer swab diameter 260 and greater than the inner swab diameter 265. The inner distance 505 can be less than the inner tube diameter 215. The inner distance 505 can be less than the plunger diameter 220.

[0057] The tube 115 can include a portion (e.g., a single prong, the plurality of prongs 120) configured to pierce the seal of the vial 105. The portion can include at least one prong. The portion comprises a first prong and a second prong separated by the inner distance 505. The portion can include the plurality of prongs 120 configured to couple with the vial 105 via an interference fit. The portion can include the plurality of prongs 120 configured to couple with the vial 105 via an interference fit. The portion can be configured to pierce the membrane seal 305. The portion can include at least one beveled prong configured to pierce the seal of the vial 105.

[0058] FIG. 6 illustrates a perspective view of the tube 115. The tube 115 can include the aperture 210, the plurality of first facets 405, and plurality of second facets 410. The aperture 210 can have an aperture diameter 605. The aperture diameter 605 can be small enough to draw the sample into the aperture 210. The aperture diameter 605 can be small enough to draw the sample into the tube 115. The aperture diameter 605 can be small enough to resist sample expulsion (e.g., sample leaving the aperture 210). The aperture diameter 605 can be determined by the pressure exerted by the swab being drawn through the plurality of prongs 120, the capillary pressure, and/or the wetting of the sample on the plurality of prongs 120. The aperture diameter 605 can be determined by the material of the plurality of prongs 120, the sample composition, and/or the formulation of the reagent 110.

[0059] Each of the plurality of prongs 120 can be separated by an outer distance 610. The outer distance 610 can be greater than the inner distance 505. The outer distance 610 can be greater than the outer swab diameter 260. The outer distance 610 can be greater than the inner swab diameter 265. The outer distance 610 can be greater than, less than, or equal to the inner tube diameter 215. The outer distance 610 can be greater than, less than, or equal to the plunger diameter 220.

[0060] FIG. 7 illustrates a perspective view of the tube 115. The tube 115 can include the plurality of prongs 120. The tube 115 can be tapered. The tube 115 can include the aperture 210 (not shown). The plunger 125 (not shown) can be inserted into the tube 115. The tube 115 can be inserted into the vial 105 (not shown).

[0061] FIG. 8 illustrates a cross-sectional view of a tube 115 (e.g., as shown in FIG. 7 taken along plane B-B). The tube 115 can include the plurality of prongs 120. The tube 115 can be tapered. The tube 115 can be hollow. The tube 115 can include the aperture 210. The plunger 125 (not shown) can be inserted into the tube 115. The tube 115 can be inserted into the vial 105 (not shown).

[0062] FIG. 9 illustrates a perspective view of the plunger 125. The plunger 125 can be inserted into the tube 115. The plunger 125 can include an end that can be pressed by a user’s thumb. The plunger 125 can include an end that can contact the blister pack 205 (not shown). The plunger 125 have a cross-shaped cross-sectional area.

[0063] FIG. 10 illustrates a method 1000 of collecting and providing a sample. The sample can include saliva. The method 1000 can include providing a kit (e.g., test kit, sample collection and detection kits, etc.). The kit can include a swab, the upper device portion assembly, and the lower device portion assembly. The kit can include a sterile, flocked swab in sealed packaging. The kit can include the upper device portion assembly in sealed packaging. The kit can include the lower device portion assembly in sealed packaging. An integrated device can include the upper device portion assembly and the lower device portion assembly. The kit can include the tube. The tube can include a gap. The kit can include the blister pack.

[0064] The method 1000 can include providing the swab, at 1005. For example, providing the swab can include providing a flocked swab. Providing the swab can include providing the flocked swab for saliva sample collection. The swab can include the sample. The swab can include a saliva sampling swab to collect saliva. The swab can include a nasal mucus sampling swab to collect nasal mucus. The sterile, flocked sampling swab can be removed from its packaging.

[0065] The method 1000 can include providing the upper device portion assembly, at 1010. The upper device portion assembly can include a tube. The tube can include a plurality of prongs. The upper device portion assembly can include a blister pack containing a liquid. The upper device portion assembly can include a plunger. The upper device portion assembly can be removed from its packaging. The sterile upper device portion assembly can be removed from its packaging.

[0066] The method 1000 can include collecting the sample (e.g., saliva, nasal mucus, blood, or other bodily fluid). The method 1000 can include inserting the swab into a patient’s mouth to collect saliva. The method 1000 can include inserting the swab into the patient’s nose to collect nasal mucus. The swab can collect and contain the sample (e.g., saliva sample, nasal mucosal sample, sputum sample, etc.).

[0067] The method 1000 can include drawing the swab containing the sample through the plurality of prongs, at 1015. For example, the saliva swab can be drawn through forks in the upper device portion assembly. The method 1000 can include collecting a predetermined amount of the sample into the tube by drawing the flocked swab between the plurality of prongs. For example, the predetermined amount of the sample can include a volume in a range of about 2 pL to 15 pL (e.g., 2 pL, 3 pL, 4 pL, 5 pL, 6 pL, 7 pL, 8 pL, 9 pL, 10 pL, 11 pL, 12 pL, 13 pL, 14 pL, or 15 pL, inclusive). The method 100 can include collecting the predetermined amount of the sample into the tube by passing the swab through the gap between the plurality of prongs. The portion can include a first prong and a second prong. The portion can include at least one beveled prong configured to pierce the seal of the vial. The method 1000 can include drawing the swab through the gap. The gap can be between the first prong and the second prong. The sample can be collected by a user (e.g., patient, technician, etc.). The user can insert the swab into the mouth for a time in a range of 5 seconds to 20 seconds (e.g., 5 seconds, 10 seconds, 15 seconds, or 20 seconds, inclusive), or until the swab is soaked with the sample. The user can collect about 100 pL of the sample. The sample-soaked swab can be drawn through the forked prong at an end of the upper device portion assembly. The integrated device can collect the predetermined amount of the sample from the sample-soaked swab. For example, from the about 100 pL of the sample collected from the user’s mouth, the integrated device can collect about 2 pL to 15 pL of sample. The integrated device can collect a precise, predetermined amount of the sample. The sample can wet the plurality of prongs. The compression of the swab can force the sample into the aperture and compress the air. When the swab is removed, the compressed air can drive the sample back out of the aperture until the pressure due to air compression is balanced by the air-liquid interface nearest the blister pack, which can act in the opposite direction, toward the blister pack.

[0068] The method 1000 can include providing the lower device portion assembly, at 1020. The lower device portion assembly can include a vial containing a reagent. The lower device portion assembly can be removed from its packaging. The sterile, sealed vial containing lyophilized reagents can be removed from its packaging.

[0069] The method 1000 can include assembling the lower device portion assembly and the upper device portion assembly, at 1025. For example, the lower device portion assembly can be pressed up against the upper device portion assembly. The plurality of prongs can puncture the vial. The method 1000 can include piercing the seal of the vial with a portion of the tube. For example, the method 1000 can include piercing the vial with the plurality of prongs. The lower device portion assembly and the upper device portion assembly can be sealed when assembled. The lower device portion assembly and the upper device portion assembly can form a sealed assembly (e.g., sealed device) when assembled. The method 1000 can include coupling the plurality of prongs with the vial via an interface fit.

[0070] The method 1000 can include depressing the plunger, at 1030. For example, the plunger can be depressed (e.g., pressed, actuated, etc.) to rupture the blister pack and drive liquid into the vial. The method 1000 can include pressing the plunger into the tube to cause the blister pack to release the liquid toward the sample to direct the sample into the vial. The user can grip the tube with four fingers and press the plunger into the tube with the thumb. The method 1000 can include puncturing the blister pack by a protrusion disposed on the tube. The method 1000 can include coupling the protrusion disposed on the tube with a recess disposed on the plunger. The user can rupture the blister pack by driving the plunger into and through the tube. The liquid contained in the blister pack can dissolve or mix with/into the reagent.

[0071] The method 1000 can include flicking the sealed assembly, at 1035 and 1040. The flick motion (e.g., downward flick motion) can drive the sample and the liquid contained in the blister pack to the bottom of the vial. The sealed assembly can be flicked to ensure that the sample, the liquid contained in the blister pack, and the reagent is at one end (e.g., the bottom) of the vial. The method 1000 can include mixing the reagent, the liquid, and the sample. The sample, the liquid contained in the blister pack, and the reagent can have a vapor pressure of 0.25 atm. The sealed assembly can resist the pressure exerted when the sealed assembly is heated.

[0072] In some embodiments, the method 1000 can include inserting the sealed device into a heating tray (e.g., heating rack, heater, etc.). The method 1000 can include inserting the vial into the heating tray. The method 1000 can include inserting the sealed device into the heating tray and heating the sealed device for 30 minutes at 65°C. Heating the sealed device for a defined temperature and time period can amplify the nucleic acids of the pathogen. Amplification of the nucleic acids can result in binding and activation of a fluorescent molecule, which can be assayed by illumination with visible light and collection of a fluorescent signal.

[0073] The method 1000 can include placing the sealed device into a detection setup subsequent to heating the sealed device. The detection setup can determine whether the sample contains a pathogen (e.g., a detectable amount of the pathogen). The results of the detection setup or test can be combined with or conveyed to a smartphone-based application. The test can be performed on a rolling time sequence (e.g., once per day). If the patient receives a negative test result, the patient may not need to repeat the test for a prescribed period of time. Heating and fluorescence detection can be performed in two separate instruments. The detection setup can include a camera, LED illumination, and specific dichroics to detect different fluorophores. The detection setup can include photomultiplier tubes (PMT) or photodiode detectors. Colormetric detection can be used as a detection method.

[0074] FIG. 11 A illustrates a side view of the plurality of prongs 120 and a front view of the plurality of prongs 120. Small volumes of saliva can be transferred from the swab 225 to the integrated device 100. The plurality of prongs 120 can include a first prong and a second prong. The first prong and the second prong can be separated by the inner distance 505 (e.g., gap distance). The first prong and the second prong can have the same length or a different length. The aperture 210 can be disposed between the first prong and the second prong.

[0075] FIG. 1 IB illustrates a plot of volume of saliva transferred between the prongs vs. saliva volume in a swab. The prongs can be beveled to pierce a tube foil seal. To determine the inner distance 505, saliva transfer volume VF from a flocked swab with a flock tip outer diameter dswab from the integrated device 100 over the range 1.3 mm < d< 2.3 mm can be measured. The data may not show a strong correlation between the inner distance 505 and VF. The experiments can result in an average VF of approximately 13 - 20 uL. The inner distance 505 can be selected based on usability. The inner distance 505 can be 1.7 mm. At the inner distance 505 < 1.7 mm, it became challenging to draw the swab 225 through the plurality of prongs 120, which can be undesirable from a user standpoint and can result in greater variance in VF. At d > 1.7 mm, it can be more difficult to tell if the swab drag had been done correctly because there can be less frictional tactile feedback as the swab is drawn through the plurality of prongs. A low volume swab with Vs < 60 pL can have a higher risk of depositing a very low trace amount of saliva when d was significantly above 1.7 mm. [0076] FIG. 11C illustrates a histogram (e.g., plot) of saliva volume transferred across multiple individuals. VF can be proportional to the volume of saliva contained in the collection swab Vs. V can be proportional to the volume of saliva contained in the collection swab Vs, as shown by the plot. To get a measure of the variability in VF between individuals, individuals can collect and transfer their own saliva from swab to device (n = 3 each).

[0077] FIG. 1 ID illustrates a time lapse of a process for transferring saliva. The saliva transfer process can be captured at 3000 frames per second. As the swab 225 is drawn between the plurality of prongs 120, a combination of surface wetting, swab fiber compression, and gravity can draw saliva from the swab 225 to the integrated device 100 and the trough beneath, as shown by the series of images. Plotting the data of the individuals, a distribution in VF can be found with a peak near 10-15 pL as shown in FIG. 1 ID with an average of 16.3 pL. This volume of saliva can be sufficient to detect low pathogen counts while keeping reagent costs low.

[0078] FIG. 12A illustrates a schematic of depressions of the plunger 125. For molecular diagnostic assays, delivery of exact volumes of aqueous media to the reaction tube can be critical. To accomplish this, the plunger 125 can be designed to translate by a well- defined distance and compress the blister by an exact amount. The initial position of the plunger 125 can be held in place by a ring 1205 (e.g., positive snap ring, internal snap ring, snap ring) in the syringe (e.g., tube 115) which seats within a negative feature (e.g., recess 1210) in the plunger 125. The mirror symmetry of the negative feature can also resist removal of the plunger 125 from the tube 115. The final position of the plunger 125 can be set by contact between the underside of the plunger rim and the top rim of the tube 115. The tube 115 can include a protrusion (e.g., ring 1205). The plunger 125 can include the recess 1210. The recess 1210 can couple with the protrusion. For example, the protrusion can sit within the recess 1210. The protrusion can slide past the recess 1210.

[0079] FIG. 12B illustrates a diagram of depression of the plunger, blister compression, and expulsion of a liquid. Compression of the blister pack 205 and the corresponding expulsion of liquid is shown in the series of images in FIG. 12B. Actuation of the integrated device 100 can be achieved by grasping the integrated device 100 in a single hand and depressing the plunger 125 by a thumb. Actuation can be accompanied by a tactile snap, which can provide the user feedback of successful actuation. [0080] FIG. 12C illustrates a plot of force vs. displacement. The force to complete the compression of a blister within the integrated device 100 can include F = 39.8 N ± 10.9 N. Partial compression of the blister pack 205 can require less force than full compression. Thus, the integrated device 100 can be designed such that plunger actuation does not fully compress the blister. To measure the force profile to actuate the integrated device 100, blister device actuation measurements can be performed using a normal force transducer. The force profile can have two peaks. The first peak in the normal force can correspond to displacement of the negative feature in the plunger 125 from the positive snap feature. Following this peak, the normal force can drop but then the normal force can increase again as the plunger 125 begins to compress the blister pack 205.

[0081] The second, larger peak can be defined as the maximum force (Fb) required to actuate the blister pack 205. Device actuation during these measurements can be performed with a fixed plunger translation velocity, v = 0.1 - 0.5 mm/s. By contrast, manual actuation can be accomplished in a less controlled manner and at a significantly higher velocity. So, while this may not directly characterize the forces required when the plunger 125 is actuated manually by hand because the rate of actuation is slower with a rheometer, it can provide some measure of the profile of resistance that the plunger 125 is overcoming. That is because this force can result in an acceleration of both the plunger 125 and the user’s thumb that results in enough kinetic energy to help compress the blister dome without requiring additional effort from the user. The user can continue pressing on the plunger 125 until it bottoms out, but the user may not have to exert extra force after the initial “snap” as the plunger disc moves past the snap ring of the syringe.

[0082] FIG. 12D illustrates a plot of force vs. blister compression fraction. The distribution of B as a function of blister compression distance can be plotted given a chosen partial compression compared to full compression of a blister. There can be a squared relationship between blister compression distance (normalized by the blister height) and the force required to actuate the blister. This can indicate that a partial compression is much easier for the user that a full compression. Since the blister can be pierced in both cases, the greater force in the full compression can be coming from the structural element of the blister dome and thus would be present even if the blister were not completely filled with water.

[0083] The integrated device 100 can be tested using an RT-LAMP formulation to show that it successfully captures the accurate amount of saliva, punctures the water-filled blister, delivers the water and saliva to the vial 105, and adequately mixes the water, saliva, and lyophilized reagents in the vial 105. The integrated device 100 can be placed in a dry bath vial heater at 65° C for 30 minutes. Saliva that has been spiked with synthetic viral RNA can be used. The LAMP reagents can be lyophilized in individual PCR vials using a lyophilizer. The sample that did not have viral RNA may not show an increased fluorescence whereas the sample spiked with viral RNA can exhibit increased fluorescence.

[0084] Table 1 illustrates a comparison of results between qPCR and the integrated device. In samples 1-3, the qPCR method detected 0 viral copies/pL and the integrated device method had a negative result (e.g., did not detect any viral copies). In sample 4, the qPCR method detected 0.14 viral copies/pL and the integrated device method had a negative result. In sample 5, the qPCR method detected 13 viral copies/pL and the integrated device method had a positive result (e.g., detected viral copies). Therefore, in this experiment, the integrated device method can detect the presence of viral copies when the qPCR method detected viral copies in a range of 0.14 to 13 viral copies/pL. In sample 6, the qPCR method detected 21 viral copies/pL and the integrated device method had a positive result. In sample 7, the qPCR method detected 116 viral copies/pL and the integrated device method had a positive result. In sample 8, the qPCR method detected 390 viral copies/pL and the integrated device method had a positive result. In sample 9, the qPCR method detected 690 viral copies/pL and the integrated device method had a positive result. In sample 10, the qPCR method detected 1320 viral copies/pL and the integrated device method had a positive result.

[0085] Table 1 : Comparison of results between qPCR test and the integrated device 100 [0086] Embodiments of the subject matter and the operations described in this specification can be implemented in digital electronic circuitry, or in computer software, firmware, or hardware, including the structures disclosed in this specification and their structural equivalents, or in combinations of one or more of them. The subject matter described in this specification can be implemented as one or more computer programs, e.g., one or more circuits of computer program instructions, encoded on one or more computer storage media for execution by, or to control the operation of, data processing apparatus. Alternatively or in addition, the program instructions can be encoded on an artificially generated propagated signal, e.g., a machine-generated electrical, optical, or electromagnetic signal that is generated to encode information for transmission to suitable receiver apparatus for execution by a data processing apparatus. A computer storage medium can be, or be included in, a computer-readable storage device, a computer-readable storage substrate, a random or serial access memory array or device, or a combination of one or more of them. Moreover, while a computer storage medium is not a propagated signal, a computer storage medium can be a source or destination of computer program instructions encoded in an artificially generated propagated signal. The computer storage medium can also be, or be included in, one or more separate components or media (e.g., multiple CDs, disks, or other storage devices).

[0087] The operations described in this specification can be performed by a data processing apparatus on data stored on one or more computer-readable storage devices or received from other sources. The term “data processing apparatus” or “computing device” encompasses various apparatuses, devices, and machines for processing data, including by way of example a programmable processor, a computer, a system on a chip, or multiple ones, or combinations of the foregoing. The apparatus can include special purpose logic circuitry, e.g., an FPGA (field programmable gate array) or an ASIC (application specific integrated circuit). The apparatus can also include, in addition to hardware, code that creates an execution environment for the computer program in question, e.g., code that constitutes processor firmware, a protocol stack, a database management system, an operating system, a cross-platform runtime environment, a virtual machine, or a combination of one or more of them. The apparatus and execution environment can realize various different computing model infrastructures, such as web services, distributed computing and grid computing infrastructures. [0088] A computer program (also known as a program, software, software application, script, or code) can be written in any form of programming language, including compiled or interpreted languages, declarative or procedural languages, and it can be deployed in any form, including as a stand-alone program or as a circuit, component, subroutine, object, or other unit suitable for use in a computing environment. A computer program may, but need not, correspond to a file in a file system. A program can be stored in a portion of a file that holds other programs or data (e.g., one or more scripts stored in a markup language document), in a single file dedicated to the program in question, or in multiple coordinated files (e.g., files that store one or more circuits, subprograms, or portions of code). A computer program can be deployed to be executed on one computer or on multiple computers that are located at one site or distributed across multiple sites and interconnected by a communication network.

[0089] Processors suitable for the execution of a computer program include, by way of example, microprocessors, and any one or more processors of a digital computer. A processor can receive instructions and data from a read only memory or a random access memory or both. The elements of a computer are a processor for performing actions in accordance with instructions and one or more memory devices for storing instructions and data. A computer can include, or be operatively coupled to receive data from or transfer data to, or both, one or more mass storage devices for storing data, e.g., magnetic, magneto optical disks, or optical disks. A computer need not have such devices. Moreover, a computer can be embedded in another device, e.g., a personal digital assistant (PDA), a Global Positioning System (GPS) receiver, or a portable storage device (e.g., a universal serial bus (USB) flash drive), to name just a few. Devices suitable for storing computer program instructions and data include all forms of non-volatile memory, media and memory devices, including by way of example semiconductor memory devices, e.g., EPROM, EEPROM, and flash memory devices; magnetic disks, e.g., internal hard disks or removable disks; magneto optical disks; and CD ROM and DVD-ROM disks. The processor and the memory can be supplemented by, or incorporated in, special purpose logic circuitry.

[0090] To provide for interaction with a user, implementations of the subject matter described in this specification can be implemented on a computer having a display device, e.g., a CRT (cathode ray tube) or LCD (liquid crystal display) monitor, for displaying information to the user and a keyboard and a pointing device, e.g., a mouse or a trackball, by which the user can provide input to the computer. Other kinds of devices can be used to provide for interaction with a user as well; for example, feedback provided to the user can be any form of sensory feedback, e.g., visual feedback, auditory feedback, or tactile feedback; and input from the user can be received in any form, including acoustic, speech, or tactile input.

[0091] The implementations described herein can be implemented in any of numerous ways including, for example, using hardware, software or a combination thereof. When implemented in software, the software code can be executed on any suitable processor or collection of processors, whether provided in a single computer or distributed among multiple computers.

[0092] Also, a computer may have one or more input and output devices. These devices can be used, among other things, to present a user interface. Examples of output devices that can be used to provide a user interface include printers or display screens for visual presentation of output and speakers or other sound generating devices for audible presentation of output. Examples of input devices that can be used for a user interface include keyboards, and pointing devices, such as mice, touch pads, and digitizing tablets. As another example, a computer may receive input information through speech recognition or in other audible format.

[0093] Such computers may be interconnected by one or more networks in any suitable form, including a local area network or a wide area network, such as an enterprise network, and intelligent network (IN) or the Internet. Such networks may be based on any suitable technology and may operate according to any suitable protocol and may include wireless networks, wired networks or fiber optic networks.

[0094] A computer employed to implement at least a portion of the functionality described herein may comprise a memory, one or more processing units (also referred to herein simply as “processors”), one or more communication interfaces, one or more display units, and one or more user input devices. The memory may comprise any computer-readable media, and may store computer instructions (also referred to herein as “processor-executable instructions”) for implementing the various functionalities described herein. The processing unit(s) may be used to execute the instructions. The communication interface(s) may be coupled to a wired or wireless network, bus, or other communication means and may therefore allow the computer to transmit communications to or receive communications from other devices. The display unit(s) may be provided, for example, to allow a user to view various information in connection with execution of the instructions. The user input device(s) may be provided, for example, to allow the user to make manual adjustments, make selections, enter data or various other information, or interact in any of a variety of manners with the processor during execution of the instructions.

[0095] The various methods or processes outlined herein may be coded as software that is executable on one or more processors that employ any one of a variety of operating systems or platforms. Additionally, such software may be written using any of a number of suitable programming languages or programming or scripting tools, and also may be compiled as executable machine language code or intermediate code that is executed on a framework or virtual machine.

[0096] In this respect, various inventive concepts may be embodied as a computer readable storage medium (or multiple computer readable storage media) (e.g., a computer memory, one or more floppy discs, compact discs, optical discs, magnetic tapes, flash memories, circuit configurations in Field Programmable Gate Arrays or other semiconductor devices, or other non-transitory medium or tangible computer storage medium) encoded with one or more programs that, when executed on one or more computers or other processors, perform methods that implement the various embodiments of the solution discussed above. The computer readable medium or media can be transportable, such that the program or programs stored thereon can be loaded onto one or more different computers or other processors to implement various aspects of the present solution as discussed above.

[0097] The terms “program” or “software” are used herein to refer to any type of computer code or set of computer-executable instructions that can be employed to program a computer or other processor to implement various aspects of embodiments as discussed above. One or more computer programs that when executed perform methods of the present solution need not reside on a single computer or processor, but may be distributed in a modular fashion amongst a number of different computers or processors to implement various aspects of the present solution.

[0098] Computer-executable instructions may be in many forms, such as program modules, executed by one or more computers or other devices. Program modules can include routines, programs, objects, components, data structures, or other components that perform particular tasks or implement particular abstract data types. The functionality of the program modules can be combined or distributed as desired in various embodiments. [0099] Also, data structures may be stored in computer-readable media in any suitable form. For simplicity of illustration, data structures may be shown to have fields that are related through location in the data structure. Such relationships may likewise be achieved by assigning storage for the fields with locations in a computer-readable medium that convey relationship between the fields. However, any suitable mechanism may be used to establish a relationship between information in fields of a data structure, including through the use of pointers, tags or other mechanisms that establish relationship between data elements.

[0100] Any references to implementations or elements or acts of the systems and methods herein referred to in the singular can include implementations including a plurality of these elements, and any references in plural to any implementation or element or act herein can include implementations including only a single element. References in the singular or plural form are not intended to limit the presently disclosed systems or methods, their components, acts, or elements to single or plural configurations. References to any act or element being based on any information, act or element may include implementations where the act or element is based at least in part on any information, act, or element.

[0101] Any implementation disclosed herein may be combined with any other implementation, and references to “an implementation,” “some implementations,” “an alternate implementation,” “various implementations,” “one implementation” or the like are not necessarily mutually exclusive and are intended to indicate that a particular feature, structure, or characteristic described in connection with the implementation may be included in at least one implementation. Such terms as used herein are not necessarily all referring to the same implementation. Any implementation may be combined with any other implementation, inclusively or exclusively, in any manner consistent with the aspects and implementations disclosed herein.

[0102] References to “or” may be construed as inclusive so that any terms described using “or” may indicate any of a single, more than one, and all of the described terms. References to at least one of a conjunctive list of terms may be construed as an inclusive OR to indicate any of a single, more than one, and all of the described terms. For example, a reference to “at least one of ‘A’ and ‘B’” can include only ‘A’, only ‘B’, as well as both ‘A’ and ‘B’. Elements other than ‘A’ and ‘B’ can also be included. [0103] The systems and methods described herein may be embodied in other specific forms without departing from the characteristics thereof. The foregoing implementations are illustrative rather than limiting of the described systems and methods.

[0104] Where technical features in the drawings, detailed description or any claim are followed by reference signs, the reference signs have been included to increase the intelligibility of the drawings, detailed description, and claims. Accordingly, neither the reference signs nor their absence have any limiting effect on the scope of any claim elements.

[0105] The systems and methods described herein may be embodied in other specific forms without departing from the characteristics thereof. The foregoing implementations are illustrative rather than limiting of the described systems and methods. Scope of the systems and methods described herein is thus indicated by the appended claims, rather than the foregoing description, and changes that come within the meaning and range of equivalency of the claims are embraced therein.

Claims

WHAT IS CLAIMED IS:

1. An integrated device for controlled sampling and detection, comprising: a vial containing a reagent; a tube comprising a plurality of prongs configured to collect a predetermined amount of sample and to pierce a seal of the vial; a blister pack containing a liquid; and a plunger configured to cause the blister pack to release the liquid toward the sample to direct the sample into the vial.

2. The integrated device of claim 1, wherein the tube comprises an aperture configured to receive the predetermined amount of sample.

3. The integrated device of claim 1, wherein the reagent comprises a lyophilized loop- mediated isothermal amplification (LAMP) reagent.

4. The integrated device of claim 1, wherein the plurality of prongs are coupled with the vial via an interference fit.

5. The integrated device of claim 1, wherein: the seal comprises a membrane seal; and the plurality of prongs is configured to pierce the membrane seal.

6. The integrated device of claim 1, wherein the plurality of prongs comprises a first prong and a second prong separated by a distance.

7. The integrated device of claim 1, further comprising: an aperture disposed between a first prong and a second prong.

8. The integrated device of claim 1, wherein the tube comprises a protrusion and the plunger comprises a recess, the recess configured to couple with the protrusion.

27

9. The integrated device of claim 1, wherein the liquid, the sample, and the reagent are configured to mix.

10. An integrated device for controlled sampling and detection, comprising: a vial containing a reagent; a tube comprising an aperture configured to collect a predetermined amount of sample, the tube comprising a portion configured to pierce a seal of the vial; a blister pack containing a liquid; and a plunger configured to cause the blister pack to release the liquid toward the sample to direct the sample into the vial.

11. The integrated device of claim 10, wherein the portion comprises at least one prong.

12. The integrated device of claim 10, wherein the reagent comprises a lyophilized loop- mediated isothermal amplification (LAMP) reagent.

13. The integrated device of claim 10, wherein the portion comprises a first prong and a second prong separated by a distance.

14. The integrated device of claim 10, wherein the portion comprises a plurality of prongs configured to couple with the vial via an interference fit.

15. The integrated device of claim 10, wherein: the seal comprises a membrane seal; and the portion is configured to pierce the membrane seal.

16. The integrated device of claim 10, wherein the tube comprises a protrusion configured to puncture the blister pack.

17. The integrated device of claim 10, wherein the portion comprises at least one beveled prong configured to pierce the seal of the vial.

18. The integrated device of claim 10, wherein the tube comprises a protrusion and the plunger comprises a recess, the recess configured to couple with the protrusion.

19. The integrated device of claim 10, wherein the liquid, the sample, and the reagent are configured to mix.

20. The integrated device of claim 10, wherein the sample comprises saliva.

21. A method of collecting and providing a sample, comprising: providing a vial containing a reagent, a tube comprising a gap, a blister pack containing a liquid, a plunger, and a swab comprising the sample; collecting a predetermined amount of the sample into the tube by passing the swab through the gap; piercing a seal of the vial with a portion of the tube; and pressing the plunger into the tube to cause the blister pack to release the liquid toward the sample to direct the sample into the vial.

22. The method of claim 21, wherein the portion comprises a first prong and a second prong, the method comprising: drawing the swab through the gap, wherein the gap is between the first prong and the second prong.

23. The method of claim 21, wherein the portion comprises a plurality of prongs, the method comprising: coupling the plurality of prongs with the vial via an interface fit.

24. The method of claim 21, comprising: mixing the reagent, the liquid, and the sample.

25. The method of claim 21, comprising: puncturing, by a protrusion disposed on the tube, the blister pack.

26. The method of claim 21, comprising: coupling a protrusion disposed on the tube with a recess disposed on the plunger.

27. The method of claim 21, wherein the sample comprises saliva.

28. The method of claim 21, wherein the portion comprises at least one beveled prong configured to pierce the seal of the vial.

29. The method of claim 21, wherein the swab is a flocked swab.