Lateral flow system and assay

• US 8,003,407 B2
• Filed: 07/29/2005
• Issued: 08/23/2011
• Est. Priority Date: 07/29/2004
• Status: Active Grant

• Current Assignee: Relia Biotechnologies (Shenzhen) Ltd.
• Original Assignee: Relia Diagnostic Systems Inc.
• Inventors: Liu, Ning, Rutter, William, Zhou, Siliang
• Primary Examiner(s): CHIN, CHRISTOPHER L
• Application Number:
  US11/572,568
• Publication Number:
  US 20090253119A1
• Time in Patent Office: 2,216 Days
• Field of Search: None
• US Class Current: 436/514
• 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 435/805 Test papers
• 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 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick

Abstract

The present invention relates to a lateral flow assay and system, including a test strip, for detection and quantification of analytes in samples, such as samples containing cells and fluid. In general, according to the present invention, a test strip for a lateral flow assay for detection of at least one analyte in a sample comprises: (1) a chromatographic strip, a sample filter, a fluid-impermeable barrier, and means for providing a mobilizable detectable agent that is capable of binding to the at least one analyte or to the capture agent after capturing the analyte to the chromatographic strip such that the mobilizable detectable agent migrates through the chromatographic strip and contacts sample that has passed through the sample filter and also has migrated through the chromatographic strip. The test strip allows detection with or without quantitation of an analyte in a sample containing whole cells.

Claims

1. A test strip for conducting a lateral flow assay for detection of at least one analyte in a sample, comprising:

(a) a chromatographic strip comprising a first end and a second end;

(b) at least one capture band, wherein the one capture band is a first capture band, and is located on the chromatographic strip, and wherein the first capture band comprises a first immobilized capture agent for capturing a first analyte in the sample;

(c) optionally, a second capture band that is located on the chromatographic strip, wherein the second capture band comprises a second immobilized capture agent for capturing a second analyte in the sample;

(d) at least one control band optionally comprising an immobilized control agent;

(e) at least one sample filter, wherein the one sample filter is a first sample filter, and wherein the first sample filter is in capillary contact with the chromatographic strip, and optionally comprises an agglutinating agent;

(f) a fluid-impermeable barrier, wherein the barrier is in capillary contact with at least the first sample filter and the chromatographic strip and slows fluid flow from the first sample filter to the chromatographic strip;

(g) at least one mobilizable detectable agent, wherein the one mobilizable detectable agent is a first mobilizable detectable agent that is capable of binding to the first analyte or to a first capture agent or to a complex formed by the combination of the first analyte and the first capture agent, wherein the first mobilizable detectable agent is provided on the chromatographic strip or can be released onto the chromatographic strip;

(h) optionally, a backing that supports the test strip;

(i) a first absorbent pad, that is in capillary contact with the chromatographic strip;

(j) optionally, a second absorbent pad that is in capillary contact with the first absorbent pad;

wherein the test strip allows detection with or without quantitation of an analyte in a sample containing whole cells.

2. The test strip of claim 1, wherein the mobilizable detectable agent is provided in a conjugate pad that is in capillary contact with the chromatographic strip.

3. The test strip of claim 2, wherein the conjugate pad retains the mobilizable detectable agent until fluid is added to the conjugate pad to release the mobilizable detectable agent.

4. The test strip of claim 1, further comprising a conjugate pad that is in capillary contact with the chromatographic strip;

a second sample filter that is in capillary contact with the chromatographic strip and optionally comprises an agglutinating agent; and

a fluid collector, wherein the fluid collector is in capillary contact or direct physical contact with (a) the second sample filter and the conjugate pad or (b) the second sample filter and the chromatographic strip.

5. The test strip of claim 1, further comprising a buffer pad that is located at or near the second end of the chromatographic strip.

6. The test strip of claim 1, further comprising a sample filter that is located at or near the second end of the chromatographic strip.

7. The test strip of claim 1, wherein the first absorbent pad is located at or near the first end of the chromatographic strip.

8. The test strip of claim 1, further comprising a conjugate pad that is located at or near the second end of the chromatographic strip.

9. The test strip of claim 1, wherein the fluid-impermeable barrier comprises a tape, optionally, a double-sided tape.

10. The test strip of claim 1, wherein the fluid-impermeable barrier is (a) completely under the first sample filter or (b) is partially under the first sample filter and is extended in the direction of the first end.

11. The test strip of claim 1, wherein the fluid-impermeable barrier is about 5 mm wide, about 5 mm to about 10 mm long and about 2.5 mm thick.

12. The test strip of claim 1, wherein the fluid-impermeable barrier is a double-sided tape made of polyester and is optionally coated with adhesive on one or both sides.

13. The test strip of claim 12, wherein the adhesive, if present, is an inert, non-migratory acrylic adhesive.

14. The test strip of claim 1, further comprising a conjugate pad, and a buffer pad, wherein the conjugate pad and the buffer pad are both at, or near, the second end, and the first sample filter, the first absorbent pad and the second absorbent pad, if present, are at, or near, the first end.

15. The test strip of claim 1, wherein the test strip further comprises a second sample filter, a fluid collector and a conjugate pad, and wherein:

(a) each of the first and second sample filters optionally comprises an agglutinating agent, and each of the first and second sample filters is in capillary contact with the chromatographic strip, the first sample filter being located at, or near, the first end of the chromatographic strip and the second sample filter being located near the second end of the chromatographic strip;

(b) the fluid collector is located between the second sample filter and the chromatographic strip, and is in capillary contact with both the second sample filter and the chromatographic strip;

(c) the conjugate pad is located at, or near, the second end of the chromatographic strip and is in capillary contact with the second sample filter and the fluid collector;

(d) the fluid-impermeable barrier is in direct physical contact with the first end of the chromatographic strip and is located under the first sample filter, where fluid flow from the first sample filter in the direction of the first end of the chromatographic strip is substantially slowed by forcing the fluid to flow underneath the impermeable barrier to reach the first end of the strip;

(e) the first absorbent pad is located at, or near, the first end of the chromatographic strip and is in direct physical contact with the chromatographic strip, wherein, the first absorbent pad is located closer to the first end of the chromatographic strip than the first sample filter; and

(f) the second absorbent pad, if present, is in capillary contact with the first absorbent pad or the chromatographic strip.

16. The test strip of claim 1, wherein the test strip further comprises a second sample filter that is in capillary contact with the chromatographic strip, a fluid collector, and a conjugate pad, and wherein:

(a) the fluid collector is located between the second sample filter and the chromatographic strip, and is in direct physical contact with both the second sample filter and the chromatographic strip;

(b) the conjugate pad is located at the second end of the chromatographic strip and is in direct physical contact with the second sample filter and indirect contact with the fluid collector; and

(c) the fluid-impermeable barrier is in direct physical contact with the first sample filter and the chromatographic strip at, or near, the first end, wherein fluid flow from the first sample filter in the direction of the first end of the chromatographic strip is substantially slowed by forcing the fluid to flow underneath the impermeable barrier to reach the first end of the strip.

21. The test strip of claim 1, wherein the mobilizable detectable agent comprises an antibody, an antigen, colored material, a label or a particle.

22. The test strip of claim 21, wherein the label comprises a colloidal particle label, a fluorescent label, a chemiluminescent label, a bioluminescent label, a redox label, a radiofrequency label, a quantum dot conjugate;

an enzyme label, a radioactive label, or a combination thereof.

23. The test strip of claim 22, wherein the colloidal particle label comprises colloidal gold particles, colloidal sulfur particles, colloidal selenium particles, colloidal barium sulfate particles, colloidal iron sulfate particles, colloidal metal iodate particles, colloidal silver halide particles, colloidal silica particles, or colloidal metal (hydrous) oxide particles.

24. The test strip of claim 22, wherein the colloidal particle label comprises a colloidal gold label.

25. The test strip of claim 21, wherein the mobilizable detectable agent is coupled to an analyte nonspecific agent, optionally, a control binding agent.

26. The test strip of claim 1, wherein the chromatographic strip comprises a nitrocellulose element.

27. The test strip of claim 1, wherein the first analyte or the second analyte, if present, comprises an antigen, an antibody, a hormone, a drug, a cell protein, a DNA, a cardiac marker, a tumor marker, a ligand, a receptor or an autoimmune disease marker.

28. The test strip of claim 1, wherein the chromatographic strip comprises at least two control bands.

29. The test strip of claim 1, comprising a backing, wherein the backing comprises a liquid impermeable material.

30. The test strip of claim 1, wherein the chromatographic strip comprises a high capacity protein binding element.

31. The test strip of claim 1, wherein:

(a) the sample contains two analytes and the chromatographic strip comprises two capture bands, each capture band comprising an immobilized capture agent that is specific for capturing one analyte but not the other, or

(b) the sample contains three analytes and the chromatographic strip comprises three capture bands, each capture band comprising an immobilized capture agent that is specific for capturing one analyte but not the other two.

32. A cassette comprising the test strip of claim 1.

33. The cassette of claim 32, wherein the cassette can be read in a device, optionally, in a device that measures reflectance.

34. The cassette of claim 32, wherein the cassette comprises a Port-1 and a Port-2 for application of sample and/or buffer.

35. A method of conducting a lateral flow assay for detection or determination of an analyte in a sample containing a fluid comprising the steps of:

(a) applying a first aliquot of the sample to the sample filter of the test strip of claim 1;

(b) causing the fluid from the sample to migrate to the first capture band;

(c) causing the first detectable agent to migrate to the first capture band;

(d) causing the first detectable agent, the analyte from the fluid from the first aliquot and the first capture band to interact; and

(e) detecting or determining the first detectable agent at the first capture band.

36. The method of claim 35, wherein the detection or determination of the first detectable agent is correlated with an amount of the analyte.

37. The method of claim 35, wherein the first detectable agent is detected or determined by an optical reflectance measurement, by fluorescence or by chemiluminescence.

38. The method of claim 35, wherein the sample contains blood cells.

39. The method of claim 35, further comprising the step of detecting or determining a second or a third detectable agent at a second or a third capture band.

17. A test strip for conducting a lateral flow assay for detection of at least one analyte in a sample, comprising:

(a) a chromatographic strip comprising a first end and a second end, at least one capture band comprising an immobilized capture agent for capturing the at least one analyte, and at least one control band comprising an immobilized control agent;

(b) a conjugate pad, wherein the conjugate pad is in capillary contact with the second end of the chromatographic strip, and wherein the conjugate pad comprises a mobilizable detectable agent that is capable of binding to the at least one analyte or to the capture agent after capturing the analyte;

(c) a fluid-impermeable barrier in direct contact with the first end of the chromatographic strip;

(d) a sample filter, wherein the sample filter is in capillary contact with the first end of the chromatographic strip and is in direct contact with the fluid-impermeable barrier such that flow from the sample filter in the direction of the first end of the chromatographic strip is substantially delayed, and wherein the sample filter optionally comprises an agglutinating agent;

(e) optionally, a buffer pad that is located at the second end of the chromatographic strip and is in direct contact with the conjugate pad;

(f) a first absorbent pad located at the first end of the chromatographic strip that is in direct contact with the chromatographic strip; and

(g) optionally, a second absorbent pad that is in direct contact with the first absorbent pad;

wherein the test strip allows detection with or without quantitation of an analyte in a sample containing whole cells.

18. A test strip for conducting a lateral flow assay for detection of at least one analyte in a sample, comprising:

(a) a chromatographic strip comprising a first end and a second end, at least one capture band comprising an immobilized capture agent for capturing the at least one analyte, and at least one control band comprising an immobilized control agent;

(b) a conjugate pad, wherein the conjugate pad is in direct capillary contact with the second end of the chromatographic strip, and wherein the conjugate pad comprises a mobilizable detectable agent that is capable of binding to the at least one analyte or to the capture agent after capturing the analyte;

(c) first and second sample filters, wherein each of the first and second sample filters optionally comprises an agglutinating agent, and each of the first and second sample filters is in capillary contact with the chromatographic strip, the first sample filter being located at, or near, the first end of the chromatographic strip and the second sample filter being located adjacent to the second end of the chromatographic strip;

(d) a fluid collector that is located between the second sample filter and the conjugate pad such that it is in direct contact with both the second sample filter and the conjugate pad;

(e) a fluid-impermeable barrier in direct contact with the first sample filter and the chromatographic strip at, or near, the first end such that flow from the first sample filter in the direction of the first end of the chromatographic strip is substantially delayed;

(f) a first absorbent pad located at the first end of the chromatographic strip that is in direct contact with the chromatographic strip, the first absorbent pad being located closer to the first end of the chromatographic strip than the first sample filter; and

(g) optionally, a second absorbent pad that is in direct contact with the first absorbent pad;

wherein the test strip allows detection with or without quantitation of an analyte in a sample containing whole cells.

19. A test strip for conducting a lateral flow assay for detection of at least one analyte in a sample, comprising:

(a) a chromatographic strip comprising a first end and a second end, at least one capture band comprising an immobilized capture agent for capturing the at least one analyte, and at least one control band comprising an immobilized control agent;

(b) first and second sample filters, wherein each of the first and second sample filters optionally comprises an agglutinating agent, and each of the first and second sample filters is in capillary contact with the chromatographic strip, the first sample filter being located at, or near, the first end of the chromatographic strip and the second sample filter being located adjacent to the second end of the chromatographic strip;

(c) a fluid collector that is located between the second sample filter and the chromatographic strip such that it is in direct contact with both the second sample filter and the chromatographic strip;

(d) a conjugate pad located at the second end of the chromatographic strip and that is in direct contact with the second sample filter and indirect contact with the fluid collector, and wherein the conjugate pad comprises a mobilizable detectable agent that is capable of binding to the at least one analyte or to the capture agent after capturing the analyte;

(e) a fluid-impermeable barrier in direct contact with the first sample filter and the chromatographic strip at, or near, the first end such that flow from the first sample filter in the direction of the first end of the chromatographic strip is substantially delayed;

(f) a first absorbent pad located at the first end of the chromatographic strip that is in direct contact with the chromatographic strip, the first absorbent pad being located closer to the first end of the chromatographic strip than the first sample filter; and

(g) optionally, a second absorbent pad that is in direct contact with the first absorbent pad;

wherein the test strip allows detection with or without quantitation of an analyte in a sample containing whole cells.

20. The test strip of claim 19, wherein each of the conjugate pad, second sample filter, and fluid collector can be offset so that the conjugate pad partially overlaps the second sample filter and the second sample filter partially overlaps the fluid collector, or the conjugate pad can partially overlap the second sample filter, and the second sample filter substantially overlaps the fluid collector.