Test strip for a lateral flow assay for a sample containing whole cells

• US 7,605,004 B2
• Filed: 07/18/2002
• Issued: 10/20/2009
• Est. Priority Date: 07/18/2001
• Status: Active Grant

• Current Assignee: Relia Biotechnologies (Shenzhen) Ltd.
• Original Assignee: Relia Diagnostic Systems Inc.
• Inventors: Zhou, Siliang
• Primary Examiner(s): Nguyen; Bao-Thuy L
• Assistant Examiner(s): DIRAMIO, JACQUELINE A
• Application Number:
  US10/200,058
• Publication Number:
  US 20030032196A1
• Time in Patent Office: 2,651 Days
• Field of Search: 422/55, 422/56, 422/57, 422/68.1, 422/101, 435/4, 435/7.1, 435/970, 4352871-2872, 435/287.7, 435/287.8, 435/287.9, 435/288.3, 435/288.7, 436/501, 436/518, 436/164, 436/169, 436/177, 436/178
• US Class Current: 436/518
• CPC Class Codes: G01N 33/525 Multi-layer analytical elements
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10T 436/25375 Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/255 including use of a solid sorbent, semipermeable membrane, or liquid extraction

Abstract

A test strip for conducting a lateral flow assay for detection of an analyte in a sample that contains cells and fluid is provided, as well as methods for making and using the test strip. This test strip is commercially usable, for example, in a diagnostic device at a point of care for detection of analytes in whole blood.

Claims

1. A test strip comprising

(a) a first membrane comprising a chromatographic strip having a reaction region, a first position, a second position and a third position;

(b) a first sample filter at the first position and having an upper surface and a lower surface, wherein the lower surface of the sample filter is in capillary contact with the chromatographic strip;

(c) a conjugate pad at the second position and having an upper surface and a lower surface, wherein the lower surface of the conjugate pad is in capillary contact with the chromatographic strip; and

(d) an absorbent pad at the third position wherein the absorbent pad is in capillary contact with the chromatographic strip when a sample is added to the sample filter, wherein the sample filter is capable of separating cells in the sample from fluid in the sample and allows the fluid to pass through to the chromatographic strip, wherein the first and second positions are on opposite sides of the reaction region, wherein the test strip facilitates lateral flow of fluid from the sample filter towards the reaction region and from the conjugate pad towards the reaction region, and wherein (i) the first position and the third position are on opposite sides of the second position, or (ii) the second position and the third position are on opposite sides of the first position.

2. The test strip of claim 1, wherein the sample filter comprises pores of varying sizes and wherein cells in the sample are retained by the sample filter and fluid in the sample passes through the sample filter.

3. The test strip of claim 2, wherein the sample filter comprises first pores having a first pore size and second pores having a second pore size, wherein the first pores are larger than the second pores and further wherein the first pores are closer to the upper surface of the sample filter and the second pores are closer to the lower surface of the sample filter.

4. The test strip of claim 1, further comprising one or more filters or membranes positioned between the sample filter and the chromatographic strip.

5. The test strip of claim 1, further comprising

(e) a reagent pad comprising an upper surface and a lower surface, wherein the lower surface of the reagent pad is in capillary contact with the upper surface of the conjugate pad.

6. The test strip of claim 5, wherein the reagent pad further comprises a reagent.

7. The test strip of claim 6, wherein the reagent is a buffer.

8. The test strip of claim 1, wherein the conjugate pad comprises a conjugate that can be released from the conjugate pad upon application of fluid to the conjugate pad, wherein the conjugate is capable of combining with an analyte to be detected to form a detectable signal.

9. The test strip of claim 1, wherein the reaction region comprises a first binding agent that is capable of binding to an analyte.

10. The test strip of claim 9, wherein the first binding agent is an antibody or antigen.

11. The test strip of claim 9, wherein the conjugate pad comprises a conjugate, and wherein the conjugate comprises a second binding agent.

12. The test strip of claim 11, wherein the second binding agent comprises a detectable marker that binds either to the first binding agent or to the analyte.

13. The test strip of claim 12, wherein the detectable marker is quantifiable.

14. The test strip of claim 8, wherein the conjugate flows from the conjugate pad to come into contact with the fluid from the sample and forms an immobilized complex.

15. The test strip of claim 5, wherein the reagent pad is a second sample filter, and the flow of fluid from the second sample filter to the conjugate pad releases a conjugate from the conjugate pad, wherein the conjugate is capable of combining with an analyte to be detected to form a detectable signal.

16. The test strip of claim 1, wherein the chromatographic strip further comprises an adhesive layer and a backing for the adhesive layer.

17. The test strip of claim 1, wherein the membrane comprises nitrocellulose.

18. The test strip of claim 1, wherein the sample filter is a blood cell/plasma separation membrane.

19. The test strip of claim 1, wherein the sample comprises fluid and cells.

20. The test strip of claim 1, wherein the sample is a blood sample, a saliva sample, a urine sample, sample taken from lymphatic tissues, a sample from a spinal tap, or a sample taken from a peritoneal cavity, synovial fluid from a joint, or fluid from lungs.

21. The test strip of claim 1, wherein the sample comprises red blood cells.

22. The test strip of claim 1, wherein the sample filter comprises a hydrophilic surface that contacts the sample.

23. The test strip of claim 1, wherein the sample filter comprises an S/G membrane.

24. The test strip of claim 3, wherein the sample filter is composed of one or more hydrophobic materials.

25. A test strip comprising

(a) a chromatographic strip having a reaction region, a first position and second position;

(b) a selective membrane;

(c) a sample filter at the first position and having an upper surface and lower surface;

(d) a conjugate pad at the second position and having an upper surface and a lower surface, wherein the lower surface of the conjugate pad is in capillary contact with the chromatographic strip; and

(e) an absorbent pad at a third position,

wherein the absorbent pad is in capillary contact with the chromatographic strip when a sample is added to the sample filter, wherein the sample filter is capable of separating cells in the sample from fluid in the sample and allows the fluid to pass through to the chromatographic strip, wherein the first and second positions are on opposite sides of the reaction region, wherein the selective membrane is in capillary contact with the sample filter and the chromatographic strip, wherein the test strip facilitates lateral flow of fluid from the selective membrane towards the reaction region and from the conjugate pad towards the reaction region, and wherein (i) the first position and the third position are on opposite sides of the second position, or (ii) the second position and the third position are on opposite sides of the first position.

26. A test strip comprising:

(a) a chromatographic strip having a reaction region;

(b) a sample filter having an upper surface and lower surface;

(c) a sample pad in capillary contact with the chromatographic strip and the lower surface of the sample filter;

(d) a conjugate pad having an upper surface and a lower surface, wherein the lower surface of the conjugate pad is in capillary contact with the chromatographic strip; and

(e) an absorbent pad,

wherein the absorbent pad is in capillary contact with the chromatographic strip when a sample is added to the sample filter, wherein the sample filter is capable of separating cells in the sample from fluid in the sample and allows the fluid to pass through to the chromatographic strip, wherein the conjugate pad and the sample pad are on opposite sides of the reaction region, wherein the test strip facilitates lateral flow of fluid from the sample pad towards the reaction region, and wherein (i) the sample filter and the absorbent pad are on opposite sides of the conjugate pad, or (ii) the conjugate pad and the absorbent pad are on opposite sides of the sample filter.

27. A test strip comprising:

(a) a chromatographic strip having a reaction region, a first position and second position;

(b) a sample filter at the first position and having an upper surface and lower surface;

(c) a sample pad having an upper surface and a lower surface, wherein the sample pad is in capillary contact with the chromatographic strip and the upper surface of the sample pad is in capillary contact with the lower surface of the sample filter;

(d) a conjugate pad at the second position and having an upper surface and a lower surface, wherein the lower surface of the conjugate pad is in capillary contact with the chromatographic strip; and

(e) an absorbent pad at a third position,

wherein the absorbent pad is in capillary contact with the chromatographic strip when a sample is added to the sample filter, wherein the sample filter is capable of separating cells in the sample from fluid in the sample and allows the fluid to pass through to the chromatographic strip, wherein the first and second positions are on opposite sides of the reaction region, wherein the test strip facilitates lateral flow of fluid from the sample pad towards the reaction region and from the conjugate pad towards the reaction region, and wherein (i) the first position and the third position are on opposite sides of the second position, or (ii) the second position and the third position are on opposite sides of the first position.

28. The test strip of claim 27, wherein the lower surface of the sample pad is in capillary contact with the chromatography strip.

29. A test strip comprising

(a) a chromatographic strip having a reaction region, a first position and second position;

(b) a sample filter at the first position and having an upper surface and lower surface;

(c) a selective membrane and a sample pad, wherein the selective membrane is either;

(i) in capillary contact with the sample filter and the sample pad, or

(ii) in capillary contact with the sample pad and the chromatographic strip;

and wherein the sample pad has an upper surface and a lower surface, wherein the sample pad is

(i) in capillary contact with the chromatographic strip if the selective membrane is in capillary contact with the sample filter and the sample pad, or

(ii) in capillary contact with the lower surface of the sample filter if the selective membrane is in capillary contact with the sample pad and the chromatographic strip;

(d) a conjugate pad at the second position and having an upper surface and a lower surface, wherein the lower surface of the conjugate pad is in capillary contact with the chromatographic strip; and

(e) an absorbent pad at a third position,

wherein the absorbent pad is in capillary contact with the chromatographic strip when a sample is added to the sample filter, wherein the sample filter is capable of separating cells in the sample from fluid in the sample and allows the fluid to pass through to the chromatographic strip, wherein the first and second positions are on opposite sides of the reaction region, wherein the test strip facilitates lateral flow of fluid from the first position towards the reaction region and from the conjugate pad towards the reaction region, and wherein (i) the first position and the third position are on opposite sides of the second position, or (ii) the second position and the third position are on opposite sides of the first position.

30. The test strip of claim 29, wherein the selective membrane is positioned under the sample filter.

31. The test strip of claim 29, wherein the selective membrane contains an agent that selectively binds to a component in the sample and removes the component from the assay environment.

32. The test strip of claim 31, wherein the agent is selected from the group consisting of an antibody, an antigen, a metabolite, a hormone, an enzyme, a virus, a prion, or parts thereof, and an agent that binds to such.