Qualitative and quantitative point-of-care assays
Disclosed herein are “equipment-free” flow-through assay devices based on patterned porous media, methods of making same, and methods of using same. The porous, hydrophilic media are patterned with hydrophobic barriers for performing assays on liquids.
1. A capillary-driven device, comprising a hydrophobic detection reagent in a porous media, wherein the hydrophobic reagent changes to hydrophilic by the presence of a target analyte, and wherein the hydrophobic detection reagent is a compound of formula (I),
wherein,
R 1 and R 2 are each independently, at each occurrence, selected from the group consisting of hydrogen, C 1 -C 6 -alkyl and C 1 -C 6 -alkoxy;
x and y are each an integer independently selected from 0, 1, 2, 3, and 4; and
n is an integer selected from 0 to 20.
2. The device of claim 1 , wherein the device is a capillary-driven vertical flow-through device, a capillary-driven lateral flow-through device, or a combination thereof.
3. The device of claim 1 , wherein the hydrophobic detection reagent is an oligomer or polymer.
4. The device of claim 1 , wherein the hydrophobic detection reagent is selected from a carbamate, ether, polyether, a poly(phthalaldehyde), a polyvinyl carbamate, a polybenzyl carbamate, or a combination thereof.
5. The device of claim 1 , wherein the hydrophobic detection reagent responds to hydrogen peroxide to convert from hydrophobic to hydrophilic molecules.
6. The device of claim 1 , wherein R 1 and R 2 are each C 1 -C 6 -alkoxy.
7. The device of claim 1 , wherein R 1 and R 2 are each methoxy.
8. The device of claim 1 , wherein n is an integer selected from 0 to 20.
9. The device of claim 1 , wherein an assay region of the device comprises one or more responsive agents that select for specific analytes.
10. The device of claim 1 , comprising an assay region derivatized with a substrate.
11. The device of claim 1 , comprising an immobilized enzyme.
12. The device of claim 1 , comprising
a first lateral flow channel leading to a first vertical flow column containing a plurality of treated layers; and
a second lateral flow channel leading to a second vertical flow column containing a plurality of treated layers.
13. The device of claim 12 , wherein the second vertical flow column is configured to correct for internal and/or external factors that affect the rate of the sample within the device, except for the effect of the target analyte on the rate.
14. The device of claim 1 , comprising a layer containing a dye for visualization of the sample in a visualization layer, wherein the dye in the dye layer is separate from the hydrophobic detection reagent.
15. The device of claim 1 , wherein the concentration of an analyte in a sample is determined by measuring the time for the sample to flow through the device after addition of the sample.
16. The device of claim 1 , wherein the concentration of an analyte in a sample is determined by visually counting the number of analyte-dependent bars after addition of the sample.