Bidirectional lateral flow test strip and method

• US 6,528,323 B1
• Filed: 06/14/1999
• Issued: 03/04/2003
• Est. Priority Date: 06/14/1999
• Status: Expired due to Term

• Current Assignee: Relia Biotechnologies (Shenzhen) Ltd.
• Original Assignee: PraxSys BioSystems, LLC
• Inventors: Sierra, George H., Dinello, Robert K., Wieck, Henry J., Thayer, Richard M., Polito, Alan J.
• Primary Examiner(s): NGUYEN, BAO THUY L
• Application Number:
  US09/332,933
• Time in Patent Office: 1,359 Days
• Field of Search: 435/7.1, 435/5, 435/7.22, 435/7.21, 435/7.31, 435/7.32, 435/7.92, 435/287.1, 435/287.2, 435/287.6, 435/287.7, 435/287.9, 435/810, 435/970, 435/974, 435/975, 436/164, 436/165, 436/169, 436/514, 436/518, 436/528, 436/530, 436/805, 436/807, 436/808, 436/810, 436/811, 436/817, 422/55-58, 422/61
• US Class Current: 436/518
• CPC Class Codes: G01N 33/54386 Analytical elements
• 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
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 435/974 Aids related test
• CPC Class Codes: Y10S 435/975 Kit
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/808 Automated or kit
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick
• CPC Class Codes: Y10S 436/811 Test for named disease, body condition or organ function
• CPC Class Codes: Y10S 436/817 Steroids or hormones

Abstract

A test strip adapted to receive a sample and detect an analyte therein is provided. The test strip comprises a sample addition zone to which a sample may be added; an absorbent zone proximal to the sample addition zone; one or more test zones distal to the sample addition zone, at least one of the test zones including a first analyte binding agent immobilized therein which is capable of binding to the analyte to be detected; and a terminal sample flow zone distal to the one or more test zones, the absorbent zone being positioned relative to the sample addition zone and having an absorption capacity relative to the other zones of the test strip such that a distal diffusion front of a sample added to the sample addition zone diffuses from the sample addition zone to a distal diffusion point within the terminal sample flow zone and then reverses direction and diffuses proximal relative to the one or more test zones.

Claims

1. A test strip adapted to receive a sample and detect an analyte therein, the test strip comprising:

a sample addition zone to which a sample may be added;

an absorbent zone positioned on the test strip in a first flow direction relative to the sample addition zone, wherein the absorbent zone comprises an absorbent pad that is affixed to the test strip;

one or more test zones positioned on the test strip in a second, different flow direction relative to the sample addition zone, at least one of the test zones including a first analyte binding agent immobilized therein which is capable of binding to the analyte to be detected, the absorbent pad being in fluid communication with the sample addition zone and having an absorption capacity relative to the other zones of the test strip such that when a volume of sample within a predetermined sample volume range for the test strip is added to the sample addition zone, the sample initially diffuses from the sample addition zone both in the first flow direction to the absorbent zone and in the second flow direction past the one or more test zones and then reverses flow, and diffuses in the first flow direction past the one or more test zones toward the absorbent zone; and

a conjugate buffer addition zone positioned on the test strip in the second flow direction relative to where the sample reverses flow, such that when conjugate buffer is added to the conjugate buffer addition zone, the conjugate buffer diffuses in the first flow direction past the one or more test zones.

2. A test strip according to claim 1 wherein the predetermined sample volume range for the test strip is between about 10 and 250 μ

L.

3. A test strip according to claim 1 wherein the predetermined sample volume range for the test strip is between about 20 and 100 μ

L.

4. A test strip according to claim 1 wherein the predetermined sample volume range for the test strip is between about 30 and 50 μ

L.

5. A test strip according to claim 1 wherein the first analyte binding agent does not bind to components in the sample other than the analyte.

6. A test strip according to claim 1 wherein the first analyte binding agent is selected from the group consisting of antibodies, engineered proteins, peptides, haptens, lysates containing heterogeneous mixtures of antigens having analyte binding sites, ligands and receptors.

7. A test strip according to claim 1 wherein the test zones further include at least a first control zone with a control binding agent immobilized therein.

8. A test strip according to claim 1 wherein the test zones include a first control zone with a control binding agent immobilized therein, and a second control zone with a same control binding agent immobilized therein as the first control zone, the first and second control zones having a different amount of the control binding agent immobilized therein.

9. A test strip according to claim 1 wherein the conjugate buffer addition zone is positioned relative to the test zones such that conjugate buffer added to the conjugate buffer addition zone at the same time as sample is added to the sample addition zone reaches the sample after the sample has reversed flow and begun diffusing in the first flow direction.

10. A test strip according to claim 1 wherein the conjugate buffer addition zone is positioned relative to the test zones such that conjugate buffer added to the conjugate buffer addition zone at the same time as sample is added to the sample addition zone reaches the test zones after the sample has reversed flow and has diffused in the first flow direction past the test zones.

11. A test strip adapted to receive a sample and detect an analyte therein, the test strip comprising:

a sample addition zone to which a sample may be added;

an absorbent zone positioned on the test strip in a first flow direction relative to the sample addition zone, wherein the absorbent zone comprises an absorbent pas that is affixed to the test strip;

one or more test zones positioned on the test-strip in a second, different flow direction relative to the sample addition zone, at least one of the test zones including a first analyte binding agent immobilized therein which is capable of binding to the analyte to be detected, the absorbent pad being in fluid communication with the sample addition zone and having an absorption capacity relative to the other zones of the test strip such that when a volume of sample within a predetermined sample volume range for the test strip is added to the sample addition zone, the sample initially diffuses from the sample addition zone both in the first flow direction to the absorbent zone and in the second flow direction past the one or more test zones and then reverses flow, and diffuses in the first flow direction past the one or more test zones toward the absorbent zone;

a conjugate buffer addition zone positioned on the test strip in the second flow direction relative to where the sample reverses flow, such that when conjugate buffer is added to the conjugate buffer addition zone, the conjugate buffer diffuses in the first flow direction past the one or more test zones; and

a zone positioned on the test strip in the second flow direction relative to where the sample reverses flow, which includes a second analyte binding agent which is capable of binding to the analyte and diffusing to the one or more test zones.

12. A test strip according to claim 11 wherein the second analyte binding agent is capable of binding to components in the sample other than the analyte.

13. A test strip according to claim 11 wherein the second analyte binding agent does not bind to components in the sample other than the analyte.

14. A test strip according to claim 11 wherein the second analyte binding agent is labeled with a detectable marker.

15. A test strip according to claim 11 wherein the second analyte binding agent is attached to a particle which is capable of diffusing to the one or more test zones.

16. A test strip according to claim 11 wherein the zone containing the second analyte binding agent is positioned on the test strip in the first flow direction relative to the conjugate buffer addition zone.

17. A test strip according to claim 11 wherein the zone containing the second analyte binding agent is the conjugate buffer addition zone.

18. A test strip comprising:

a first fluid addition zone to which a first fluid may be added;

an absorbent zone positioned on the test strip in a first flow direction relative to the first fluid addition zone, wherein the absorbent zone comprises an absorbent pas that is affixed to the test strip;

one or more test zones positioned on the test strip in a second, different flow direction relative to the first fluid addition zone, at least one of the test zones including a first analyte binding agent immobilized therein which is capable of binding to an analyte to be detected using the test strip, the absorbent pad being in fluid communication with the first fluid addition zone and having an absorption capacity relative to the other zones of the test strip such that when a volume of the first fluid within a predetermined volume range for the test strip is added to the first fluid addition zone, the first fluid initially diffuses from the first fluid addition zone both in the first flow direction to the absorbent zone and in the second flow direction past the one or more test zones and then, because of absorption by the absorbent pad, first fluid diffusing in the second direction reverses flow, and diffuses in the first flow direction past the one or more test zones toward the absorbent zone; and

a second fluid addition zone positioned on the test strip in the second flow direction relative to where the first fluid reverses flow, such that when the second fluid is added to the second fluid addition zone, the second fluid diffuses in the first flow direction past the one or more test zones.

19. A test strip according to claim 18 wherein the predetermined sample volume range for the test strip is between about 10 and 250 μ

L.

20. A test strip according to claim 18 wherein the predetermined sample volume range for the test strip is between about 20 and 100 μ

L.

21. A test strip according to claim 18 wherein the predetermined sample volume range for the test strip is between about 30 and 50 μ

L.

22. A test strip according to claim 18 wherein the first analyte binding agent is selected from the group consisting of antibodies, engineered proteins, peptides, haptens, lysates containing heterogeneous mixtures of antigens having analyte binding sites, ligands and receptors.

23. A test strip according to claim 18 wherein the test zones further include at least a first control zone with a control binding agent immobilized therein.

24. A test strip according to claim 18 wherein the test zones include a first control zone with a control binding agent immobilized therein, and a second control zone with a same control binding agent immobilized therein as the first control zone, the first and second control zones having a different amount of the control binding agent immobilized therein.

25. A test strip according to claim 18 wherein the second fluid addition zone is positioned relative to the test zones such that second fluid added to the second fluid addition zone at the same time as the first fluid is added to the first fluid addition zone does not reach the first fluid before the first fluid reverses flow.

26. A test strip according to claim 18 wherein the second fluid addition zone is positioned relative to the test zones such that second fluid added to the second fluid addition zone at the same time as first fluid is added to the first fluid addition zone reaches the test zones after the first fluid reverses flow and diffuses in the first flow direction past the one or more test zones toward the absorbent zone.

27. A method for detecting an analyte in a sample comprising:

applying the sample to a test strip wherein the sample (a) diffuses in a first flow direction toward an absorbent pad and in a second, different flow direction to one or more test zones, at least one of the test zones including a first analyte binding agent immobilized therein which binds to analyte in the sample, (b) diffuses past the one or more test zones in the second flow direction, and then (c) reverses flow and diffuses in the first flow direction past the one or more test zones, towards the absorbent pad;

delivering a conjugate buffer to a conjugate buffer addition zone which is positioned on the test strip in the second flow direction relative to where the sample reversed flow, delivery of the conjugate buffer causing a second analyte binding agent to diffuse in the first flow direction to the one or more test zones after the sample has reversed flow and diffuses in the first flow direction past the one or more test zones, the second analyte binding agent binding to analyte immobilized in the test zones; and

detecting the second analyte binding agent immobilized in the test zones which indicates that analyte is present in the sample.

28. A method according to claim 27 wherein the conjugate buffer is added to the test strip at a same time as the sample is added to the test strip.

29. A method according to claim 27 wherein the conjugate buffer is added to the test strip before the sample is added to the test strip.

30. A method according to claim 27 wherein the conjugate buffer is added to the test strip after the sample is added to the test strip.

31. A method according to claim 27 wherein the first analyte binding agent does not bind to components in the sample other than the analyte.

32. A method according to claim 27 wherein the first analyte binding agent is selected from the group consisting of antibodies, engineered proteins, peptides, haptens, lysates containing heterogeneous mixtures of antigens having analyte binding sites, ligands and receptors.

33. A method according to claim 27 wherein the second analyte binding agent is contained on the test strip where the conjugate buffer is delivered, delivery of the conjugate buffer causing the diffusion of the second analyte binding agent.

34. A method according to claim 27 wherein the second analyte binding agent is contained on the test strip in a first flow direction relative to where the conjugate buffer is delivered, delivery of the conjugate buffer causing the diffusion of the second analyte binding agent.

35. A method according to claim 27 wherein delivering the conjugate buffer to the test strip includes delivering the second analyte binding agent to the test strip within the conjugate buffer.

36. A method according to claim 27 wherein the test zones include at least a first control zone with a control binding agent immobilized therein, delivering the conjugate buffer causing a control agent to diffuse to the first control zone and bind to the control binding agent immobilized therein.

37. A method according to claim 27 wherein the test zones include first and second control zones which each include a different amount of a control binding agent immobilized therein, delivering the conjugate buffer causing a control agent to diffuse to the first and second control zones and bind to the control binding agent immobilized therein.

38. A method according to claim 27 wherein the second analyte binding agent is capable of binding to components in the sample other than the analyte.

39. A method according to claim 27 wherein the second analyte binding agent does not bind to components in the sample other than the analyte.

40. A method according to claim 27 wherein the second analyte binding agent is labeled with a detectable marker, detecting the second analyte binding agent including detecting the detectable marker.

41. A method according to claim 27 wherein the second analyte binding agent is attached to a particle, detecting the second analyte binding agent including detecting the particle.