The disclosure provides low cost, portable three-dimensional devices for performing multiplexed assays. The devices comprise at least two substantially planar layers disposed in parallel planes, wherein one of the layers is movable relative to each other parallel to the planes to permit the establis
The disclosure provides low cost, portable three-dimensional devices for performing multiplexed assays. The devices comprise at least two substantially planar layers disposed in parallel planes, wherein one of the layers is movable relative to each other parallel to the planes to permit the establishment of fluid flow communication serially between the two layers.
대표청구항▼
1. A device for assay of a fluid sample, the device comprising: at least first and second substantially planar members disposed in the same or parallel planes, wherein the first substantially planar member is a porous, hydrophilic, adsorbent material comprising fluid-impermeable barriers that define
1. A device for assay of a fluid sample, the device comprising: at least first and second substantially planar members disposed in the same or parallel planes, wherein the first substantially planar member is a porous, hydrophilic, adsorbent material comprising fluid-impermeable barriers that define boundaries of plural hydrophilic regions and the second substantially planar member defines a test zone for presentation of a sample for assay;said plural hydrophilic regions and said test zone comprising porous, hydrophilic, adsorbent material for transfer of fluid within the porous, hydrophilic, adsorbent material by capillary action;said fluid-impermeable barriers penetrating the first planar member to define the boundaries of the plural hydrophilic regions through which fluid flows by capillary action;said members being moveable relative to each other to permit establishment of fluid flow communication serially between at least two of said hydrophilic regions and the test zone;a reagent disposed in said device within or in flow communication with one of said hydrophilic regions and in flow communication with said test zone when said one hydrophilic region and test zone are in fluid flow communication. 2. The device of claim 1 comprising at least two separate test zones. 3. The device of claim 2 further comprising at least two reagents disposed in said device within or in flow communication with separate said hydrophilic regions and in flow communication with respective said separate test zones when said respective hydrophilic regions and test zones are in fluid flow communication, thereby to permit execution of assays for multiple analytes substantially simultaneously. 4. The device of claim 1 further comprising a positive or a negative control zone in the member comprising the test zone. 5. The device of claim 1 further comprising in the member comprising the test zone a plurality of positive control zones comprising known concentrations of an analyte thereby to permit assessment of concentration of an analyte in a sample. 6. The device of claim 1 comprising plural reagents for treating said sample in said device within or in flow communication with one or more of said hydrophilic regions and in flow communication with said test zone when said one hydrophilic region and test zone are in fluid flow communication. 7. The device of claim 1 wherein a said reagent functions to develop color in a said test zone as an indication of the presence, absence or concentration of an analyte in a sample. 8. The device of claim 1 comprising a washing reagent within or in fluid communication with a second hydrophilic zone which washing reagent functions to wash an analyte bound to a test zone by removing unbound species therein when said second hydrophilic region and test zone are in fluid flow communication. 9. The device of claim 1 wherein establishment of fluid flow communication between a said hydrophilic region and the test zone is effected by movement of said first and second members relative to each other to register vertically or horizontally a said test zone and a said hydrophilic region. 10. The device of claim 1 further comprising a carrier fluid inlet and a series of flow paths between said carrier fluid inlet and said hydrophilic regions. 11. The device of claim 1 further comprising a sample inlet in fluid communication with a said test zone. 12. The device of claim 1 further comprising a sample filter upstream of and in fluid communication with a said test zone. 13. The device of claim 1 further comprising a reagent reservoir upstream of and in fluid communication with a said test zone comprising a reagent for pre-treating a sample. 14. The device of claim 1 wherein said test zone comprises an immobilized analyte binder. 15. The device of claim 1 comprising a blocking agent disposed in said device within or in flow communication with one of said hydrophilic regions. 16. The device of claim 1 comprising an antibody reagent disposed in said device within or in flow communication with one of said hydrophilic regions. 17. The device of claim 14 comprising a labeled antibody reagent. 18. The device of claim 17 wherein the antibody is labeled with an enzyme, a fluorophore, or a colored particle to permit colorimetric assessment of analyte presence or concentration. 19. The device of claim 1 comprising an enzyme substrate disposed in said device within or in flow communication with one of said hydrophilic regions. 20. The device of claim 18 wherein the enzyme is alkaline phosphatase or horseradish peroxidase. 21. The device of claim 19 wherein the substrate is selected from the group consisting of BCIP/NBT, 3,3′,5,5′-Tetramethylbenzidine (TMB), 3,3′-Diaminobenzidine (DAB) and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS). 22. The device of claim 1 wherein the analyte is selected from the group consisting of: viral antigens, bacterial antigens, fungal antigens, parasitic antigens, cancer antigens, and metabolic markers. 23. The device of claim 1 further comprising visual indicia of the establishment of fluid communication of a test zone with plural said hydrophilic regions. 24. The device of claim 1 wherein said members comprise a material selected from the group consisting of paper, cloth, and polymer film. 25. The device of claim 1 wherein said fluid-impermeable barriers that define boundaries of said plural hydrophilic regions are produced by screening, stamping, printing or photolithography and comprise a photoresist, wax, poly(methylmethacrylate), an acrylate polymer, polystyrene, polyethylene, polyvinylchloride, a fluoropolymer, or a photo-polymerizable polymer that forms a hydrophobic polymer. 26. The device of claim 1 further comprising a fluid-impermeable layer disposed between adjacent layers and defining openings permitting fluid flow therethrough. 27. The device of claim 1 further comprising an adsorbent layer for drawing fluid from or through a said hydrophilic region and through a said test zone. 28. An assay method comprising providing the device of claim 1, adding a sample to said test zone, moving one said layer in relation to another to establish serially fluid communication between a test zone and said hydrophilic zones to permit fluid flow therebetween for a time interval and to execute multiple steps of an assay, and examining said test zone to determine the presence, absence, or concentration of an analyte. 29. An assay method comprising providing the device of claim 1, adding a sample to said test zone, moving one said layer in relation to another to establish serially fluid communication between a test zone and said hydrophilic zones to permit fluid flow therebetween for a time interval and to execute multiple steps of an assay, and examining the development of or intensity of color development in said test zone to determine the presence, absence, or concentration of an analyte. 30. The method of claim 29 comprising the additional step of creating digital data indicative of an image of said test zone and therefore of the assay result, and transmitting the data remotely for analysis to obtain diagnostic information. 31. The device of claim 1 wherein said members comprise paper. 32. The device of claim 26 wherein said members comprise paper. 33. The device of claim 31 wherein said fluid-impermeable barriers that define boundaries of said plural hydrophilic regions comprise wax. 34. The device of claim 32 wherein said fluid-impermeable barriers that define boundaries of said plural hydrophilic regions comprise wax.
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