Lateral flow assays using two dimensional test and control signal readout patterns

• US 9,599,615 B2
• Filed: 09/12/2014
• Issued: 03/21/2017
• Est. Priority Date: 03/13/2013
• Status: Active Grant

• Current Assignee: Symbolics, LLC
• Original Assignee: Symbolics, LLC
• Inventors: O'Farrell, Brendan, Tisone, Thomas C.
• Primary Examiner(s): Chin, Chris L
• Application Number:
  US14/485,283
• Publication Number:
  US 20150080254A1
• Time in Patent Office: 921 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: B01L 2200/148 Specific details about calibrations
• CPC Class Codes: B01L 2300/028 Graduation
• CPC Class Codes: B01L 3/5023 with a sample being transported to, and subsequently stored in an absorbent for analysis
• CPC Class Codes: G01N 33/54388 based on lateral flow
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387

Abstract

The present invention relates to novel lateral flow devices using two dimensional features, preferably, uniform two dimensional test and control signal readout patterns, and the methods for detecting an analyte using the lateral flow devices.

Claims

1. A test device for detecting an analyte in a liquid sample, which device comprises a plurality of reagent dots on a matrix, wherein at least two of said reagent dots do not overlap and are sufficiently spaced apart from each other so that when said liquid sample flows laterally along said matrix, flow of said liquid sample to, through and/or around one of said two reagent dots does not substantially affect flow of said liquid sample to, through and/or around said other reagent dot, and:

1) after a liquid sample containing no analyte or an analyte below detection level flows laterally along said test device and passes said plurality of reagent dots, said plurality of reagent dots form two separate two-dimensional signal features A and B, and said feature A, feature B or a combination thereof, viewed from a direction substantially parallel or opposite to the liquid sample flow direction, indicates that a test is properly conducted and said sample contains no analyte or the analyte below detection level;

or

2) after a liquid sample containing an analyte at or above detection level flows laterally along said test device and passes said plurality of reagent dots, said plurality of reagent dots;

a) form one discernible feature that is one of said features A and B, form additional signal feature C that obscures the other of said features A and B to form an obscured feature D, and said discernible feature from features A and B or a combination of said discernible feature from features A and B and said obscured feature D, viewed from a direction substantially parallel or opposite to the liquid sample flow direction, indicates that a test is properly conducted; and

b) form at least one additional two-dimensional signal feature E, and a combination of said discernible feature from features A and B and said feature E, viewed from a direction substantially perpendicular to the liquid sample flow direction, indicates that said sample contains said analyte at or above detection level.

2. The test device of claim 1, wherein the plurality of reagent dots form at least two additional two-dimensional signal features E and E′

, and a combination of the discernible feature from features A and B and the features E and E′

, viewed from a direction substantially perpendicular to the liquid sample flow direction, indicates that the sample contains the analyte at or above detection level.

3. The test device of claim 1, wherein at least one, some or all of the features A-E′

, or a combination thereof, comprises a predetermined pattern that is selected from the group consisting of a line, multiple lines, a symbol, a geometric shape and an alpha-numeric shape.

4. The test device of claim 3, wherein the alpha-numeric shape is a letter, a word, a number or a combination thereof.

5. The test device of claim 4, wherein the combination of the features A and B, viewed from a direction substantially parallel or opposite to the liquid sample flow direction, forms the word “

NO”

.

6. The test device of claim 4, wherein the combination of the discernible feature from features A and B and the features E and E′

, viewed from a direction substantially perpendicular to the liquid sample flow direction, forms the phrase “

POS”

.

7. The test device of claim 3, wherein the obscured feature D is a geometric shape.

8. The test device of claim 1, wherein the matrix comprises, at the locations for generating features A and/or B, a control reagent.

9. The test device of claim 1, wherein the matrix comprises, at the locations for generating features C, E, and/or E and E′

, a test reagent.

17. The test device of claim 1, wherein the matrix comprises, at the locations for generating features A and/or B, a control reagent.

18. The test device of claim 1, wherein the matrix comprises, at the locations for generating features C, E, and/or E and E′

, a test reagent.

19. The test device of claim 1, wherein the plurality of reagent dots comprises at least 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 500, 1,000, 5,000, 10,000 or more reagent dots.

20. The test device of claim 1, wherein at least a quarter, a third, half or all reagent dots do not overlap and are sufficiently spaced apart from each other so that when the liquid sample flows laterally along the matrix, flow of the liquid sample to, through and/or around one of the reagent dots does not substantially affect flow of the liquid sample to, through and/or around the other reagent dots.

21. The test device of claim 1, wherein at least one of the reagent dots has a diameter of about 0.1-1 um, 1-10 um, 10-50 um, 51-100 um, 101-200 um, 201-300 um, 301-400 um, 401-500 um and 501-1000 um, or at least one of the reagent dots has a diameter or surface area that is about 10%, 5%, 1%, 0.5%, 0.1%, 0.05%, 0.01%, 0.001% or smaller diameter or surface area of the length, width or surface area of the matrix.

22. The test device of claim 1, wherein the distance between edges of at least two of the reagent dots is about 1-10 um, 10-50 um, 51-100 um, 101-200 um, 201-300 um, 301-400, 401-500, or 501-600 um.

23. A method for detecting an analyte in a liquid sample, which method comprises:

a) contacting a liquid sample with the test device of claim 1, wherein the liquid sample is applied to a site of the test device upstream of the plurality of reagent dots;

b) transporting an analyte, if present in the liquid sample, and a labeled reagent to the plurality of reagent dots; and

c) assessing the presence, absence, amount and/or pattern of signal(s) generated by the labeled reagent at the plurality of reagent dots to determining the presence, absence and/or amount of the analyte in the liquid sample.

24. The method of claim 23, wherein the liquid sample and the labeled reagent are premixed to form a mixture and the mixture is applied to the test device.

25. The method of claim 23, wherein the test device comprises a dried labeled reagent before use and the dried labeled reagent is solubilized or resuspended, and transported to the plurality of reagent dots by the liquid sample.

26. The method of claim 23, wherein the liquid sample is body fluid sample.

10. A test device for detecting an analyte in a liquid sample, which device comprises a plurality of reagent dots on a matrix, wherein at least two of said reagent dots do not overlap and are sufficiently spaced apart from each other so that when said liquid sample flows laterally along said matrix, flow of said liquid sample to, through and/or around one of said two reagent dots does not substantially affect flow of said liquid sample to, through and/or around said other reagent dot, and:

1) after a liquid sample containing no analyte or an analyte below detection level flows laterally along said test device and passes said plurality of reagent dots, said plurality of reagent dots form two separate two-dimensional signal features A and B, said feature A, feature B or a combination thereof, viewed from a direction substantially parallel or opposite to the liquid sample flow direction, indicates that a test is properly conducted and said sample contains no analyte or the analyte below detection level;

or

2) after a liquid sample containing an analyte at or above detection level flows laterally along said test device and passes said plurality of reagent dots, said plurality of reagent dots;

a) form two separate two-dimensional signal features A and B and an additional signal feature C, said feature A, feature B or a combination thereof, viewed from a direction substantially parallel or opposite to the liquid sample flow direction, indicates that a test is properly conducted, and a combination of one or both of said features A and B and said feature C, viewed from a direction substantially parallel or opposite to the liquid sample flow direction, generates a signal that aids interpretation of the test result; and

b) form at least one additional two-dimensional signal feature E, and a combination of at least one of said features A and B and said feature E, viewed from a direction substantially perpendicular to the liquid sample flow direction, indicates that said sample contains said analyte at or above detection level.

11. The test device of claim 10, wherein the plurality of reagent dots form at least two additional two-dimensional signal features E and E′

, and a combination of one of the features A and B and the features E and E′

, viewed from a direction substantially perpendicular to the liquid sample flow direction, indicates that the sample contains the analyte at or above detection level.

12. The test device of claim 11, wherein at least one, some or all of the features A-E′

, or a combination thereof, comprises a predetermined pattern that is selected from the group consisting of a line, multiple lines, a symbol, a geometric shape and an alpha-numeric shape.

13. The test device of claim 12, wherein the alpha-numeric shape is a letter, a word, a number or a combination thereof.

14. The test device of claim 13, wherein the combination of the features A and B, viewed from a direction substantially parallel or opposite to the liquid sample flow direction, forms the word “

NO”

.

15. The test device of claim 13, wherein the combination of one of the features A and B and the features E and E′

, viewed from a direction substantially perpendicular to the liquid sample flow direction, forms the phrase “

POS”

.

16. The test device of claim 13, wherein the combination of the features A, B and C forms the word “

NOW”

.