Test strip apparatus and method for determining presence of analyte in a fluid sample

• US 6,001,658 A
• Filed: 09/13/1996
• Issued: 12/14/1999
• Est. Priority Date: 09/13/1996
• Status: Expired due to Term

• Current Assignee: Sekisui Diagnostics, LLC (Sanofi )
• Original Assignee: Diagnostic Chemicals Limited
• Inventors: Fredrickson, Robert A.
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US08/710,259
• Time in Patent Office: 1,187 Days
• Field of Search: 422/56-58, 422/61, 435/7.9-7.95, 435/287.1, 435/287.2, 435/287.7, 435/287.9, 435/805, 435/810, 435/970, 436/514, 436/518, 436/525, 436/531, 436/533, 436/534, 436/169, 436/805, 436/810
• 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 436/805 Optical property
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick

Abstract

An apparatus in the form of a test strip is useful for determining the presence of one or more components in a fluid sample. The test strip can be used by itself or in conjunction with an associated housing assembly. The test strip provides a rapid volume, timing and temperature independent visually readable strip for the semi-quantitative detection of a component, such as an analyte.

Claims

1. A semiquantitative method for determining the presence of an analyte in a sample fluid, comprising

exposing a portion of a test strip apparatus to said sample fluid to be tested, wherein said test strip apparatus comprises:

a conjugate pad comprising a diffusible labelled first binding partner that binds with said analyte;

a first area comprising an immobilized second binding partner that competes with said analyte to bind with said diffusible labelled first binding partner; and

a second area comprising an immobilized third binding partner that binds with said diffusible labelled first binding partner,

located in sequence in a capillary fluid flow direction in said test strip apparatus;

allowing said analyte in said sample fluid to migrate up the test strip apparatus by capillary action; and

reading a result of said test strip by comparing relative label intensities in said first area and said second area,

wherein a greater relative label intensity in said second area as compared to said first area indicates the presence of said analyte in said sample fluid in an amount greater than a value determined by the amounts of said first and second binding partners and a lesser relative intensity in said second area as compared to said first area indicates the presence of said analyte in said sample fluid in an amount less than a value determined by the amounts of said first and second binding partners.

2. A method according to claim 1, wherein in an absence of said analyte in said sample fluid, at least 80% of said diffusible labelled first binding partner binds to said immobilized second binding partner.

3. A method according to claim 1, wherein said diffusible labelled first binding partner comprises an antibody-latex microparticle conjugate, said immobilized second binding partner comprises human serum albumin, said immobilized third binding partner comprises an anti-mouse antibody capable of binding with said antibody-latex microparticle conjugate, and said analyte is human urine albumin.

4. A method according to claim 1, wherein said test strip apparatus further comprises a sample pad located upstream of said diffusible labelled first binding partner in said capillary fluid flow direction, and a draw wick pad located downstream of said immobilized third binding partner in said capillary fluid flow direction in said test strip apparatus.

5. A method according to claim 1, wherein said immobilized second binding partner is selected from the group consisting of immobilized analyte and immobilized analyte analog.

6. A method according to claim 1, wherein in an absence of said analyte in said sample fluid, at least 95% of said diffusible labelled first binding partner binds to said immobilized second binding partner.

7. A method according to claim 1, wherein said diffusible labelled first binding partner comprises an antibody-colloidal gold conjugate.

8. A method according to claim 1, wherein said diffusible labelled first binding partner is an antibody-label conjugate.

9. A method according to claim 1, wherein said diffusible labelled first binding partner comprises a visible label.

10. A method according to claim 9, wherein said visible label comprises latex microparticles.

11. A method according to claim 1, wherein said immobilized third binding partner comprises an antibody.

12. A method according to claim 1, wherein said test strip apparatus further comprises a metering component directed against said analyte to sequester a portion of said analyte to meter a concentration of said analyte in said sample fluid, wherein said metering component is located in said test strip apparatus upstream of said immobilized second binding partner in said capillary fluid flow direction.

13. A method according to claim 12, wherein said metering component is an unlabelled antibody.

14. The method of claim 1, wherein said first area comprises sufficient immobilized second binding partner to bind 80-99% of said labelled first binding partner in an absence of said analyte in said sample fluid.

15. The method of claim 1, wherein said first area comprises sufficient immobilized second binding partner to bind 90-99% of said labelled first binding partner in an absence of said analyte in said sample fluid.

16. The method of claim 1, wherein said first area comprises sufficient immobilized second binding partner to bind about 95% of said labelled first binding partner in an absence of said analyte in said sample fluid.

17. A test strip apparatus for semiquantitative determination of an analyte in a sample fluid, comprising

a sample pad;

a conjugate pad comprising a diffusible labelled first binding partner that binds with said analyte;

a first area comprising an immobilized second binding partner that competes with said analyte to bind with said diffusible labelled first binding partner, wherein there is a sufficient amount of said second binding partner to bind at least 80%, but no more than 99%, of said labelled first binding partner in an absence of said analyte in said sample fluid;

a second area comprising an immobilized third binding partner that binds with said diffusible labelled first binding partner; and

a draw wick pad,

located in sequence in a capillary fluid flow direction in said test strip apparatus,

wherein when said test strip is exposed to said sample fluid, a greater relative label intensity in said second area as compared to said first area indicates the presence of said analyte in said sample fluid in an amount greater than a value determined by the amounts of said first and second binding partners and a lesser relative intensity in said second area as compared to said first area indicates the presence of said analyte in said sample fluid in an amount less than a value determined by the amounts of said first and second binding partners.

18. A test strip apparatus according to claim 17, wherein said diffusible labelled first binding partner comprises an antibody-latex microparticle conjugate, said immobilized second binding partner comprises human serum albumin, and said immobilized third binding partner comprises an anti-mouse antibody capable of binding with said antibody-latex microparticle conjugate.

19. The apparatus of claim 17, wherein there is a sufficient amount of said second binding partner to bind 90-99% of said first binding partner in an absence of said analyte in said sample fluid.

20. The apparatus of claim 17, wherein there is a sufficient amount of said second binding partner to bind about 95% of said first binding partner in an absence of said analyte in said sample fluid.

21. The apparatus of claim 17, wherein said immobilized second binding partner is selected from the group consisting of immobilized analyte and immobilized analyte analog.

22. A semiquantitative method for determining the presence of an analyte in a sample fluid, comprising

exposing a portion of a test strip apparatus to said sample fluid to be tested, wherein said test strip apparatus comprises:

a conjugate pad comprising a diffusible labelled first binding partner that binds with said analyte;

a first area comprising an immobilized second binding partner that binds with analyte which has bound to said diffusible labelled first binding partner to form an analyte sandwich, but does not bind with said diffusible labelled first binding partner; and

a second area comprising an immobilized third binding partner that binds with said diffusible labelled first binding partner,

located in sequence in a capillary fluid flow direction in said test strip apparatus;

allowing said analyte in said sample fluid to migrate up the test strip apparatus by capillary action; and

reading a result of said test strip by comparing relative label intensities in said first area and said second area,

wherein a greater relative label intensity in said first area as compared to said second area indicates the presence of said analyte in said sample fluid in an amount greater than a value determined by the amounts of said first, second and third binding partners and a lesser relative label intensity in said first area as compared to said second area indicates the presence of said an analyte in said sample fluid in an amount less than a value determined by the amounts of said first, second and third binding partners.

23. The method of claim 22, wherein said immobilized second binding partner is selected from the group consisting of immobilized antibody to said analyte and an antibody recognizing a different epitope on said analyte than the epitope recognized by said labelled first binding partner.

24. A semiquantitative method for determining the presence of an analyte in a sample fluid, comprising

exposing a portion of a test strip apparatus to said sample fluid to be tested, wherein said test strip apparatus comprises:

a conjugate pad comprising a diffusible labelled form of said analyte;

a first area comprising an immobilized first binding partner that binds with said analyte and said labelled form of said analyte; and

a second area comprising an immobilized second binding partner that binds with any of said analyte not bound to said immobilized first binding partner,

located in sequence in a capillary fluid flow direction in said test strip apparatus;

allowing said analyte in said sample fluid to migrate up the test strip apparatus by capillary action; and

reading a result of said test strip by comparing relative label intensities in said first area and said second area,

wherein a greater relative label intensity in said second area as compared to said first area indicates the presence of said analyte in said sample fluid in an amount greater than a value determined by the amount of said first binding partner and a lesser relative label intensity in said second area as compared to said first area indicates the presence of said analyte in said sample fluid in an amount less than a value determined by the amount of said first binding partner.

25. The method of claim 24, wherein said first area comprises sufficient immobilized first binding partner to bind 80-99% of said labelled analyte in an absence of said analyte in said sample fluid.

26. The method of claim 24, wherein said first area comprises sufficient immobilized first binding partner to bind 90-99% of said labelled analyte in an absence of said analyte in said sample fluid.

27. The method of claim 24, wherein said first area comprises sufficient immobilized first binding partner to bind about 95% of said labelled analyte in an absence of said analyte in said sample fluid.

28. The method of claim 24, wherein said immobilized first binding partner is selected from the group consisting of immobilized polyclonal antibody against said analyte and monoclonal antibody against said analyte.

29. A test strip apparatus for determining the presence of an analyte in a sample fluid, comprising

a sample pad;

a conjugate pad comprising a diffusible labelled first binding partner that binds with said analyte;

a metering component;

a first area comprising an immobilized second binding partner that competes with said analyte to bind with said diffusible labelled first binding partner;

a second area comprising an immobilized third binding partner that binds with said diffusible labelled first binding partner; and

a draw wick pad,

located in sequence in a capillary fluid flow direction in said test strip apparatus.

30. The apparatus of claim 29, wherein in an absence of said analyte in said sample fluid, at least 80% of said diffusible labelled first binding partner binds to said immobilized second binding partner in an absence of said analyte in said sample fluid.

31. The apparatus of claim 29, wherein said diffusible labelled first binding partner comprises an antibody-latex microparticle conjugate, said immobilized second binding partner comprises human serum albumin, said immobilized third binding partner comprises an anti-mouse antibody capable of binding with said antibody-latex microparticle conjugate, and said analyte is human urine albumin.

32. The apparatus of claim 29, wherein said immobilized second binding partner is selected from the group consisting of immobilized analyte and immobilized analyte analog.

33. The apparatus of claim 29, wherein in an absence of said analyte in said sample fluid, at least 95% of said diffusible labelled first binding partner binds to said immobilized second binding partner in an absence of said analyte in said sample fluid.

34. The apparatus of claim 29, wherein said diffusible labelled first binding partner comprises an antibody-colloidal gold conjugate.

35. The apparatus of claim 29, wherein said diffusible labelled first binding partner is an antibody-label conjugate.

36. The apparatus of claim 29, wherein said diffusible labelled first binding partner comprises a visible label.

37. The apparatus of claim 36, wherein said visible label comprises latex microparticles.

38. The apparatus of claim 29, wherein said immobilized third binding partner comprises an antibody.

39. The apparatus of claim 29, wherein said metering component is directed against said analyte to sequester a portion of said analyte to meter a concentration of said analyte in said sample fluid.

40. The apparatus of claim 39, wherein said metering component is an unlabelled antibody.

41. The apparatus of claim 29, wherein said metering component is contained in said conjugate pad.

42. The apparatus of claim 29, wherein said metering component is contained in said sample pad.

43. The apparatus of claim 29, wherein said metering component is contained in a metering pad located prior to said first area in said capillary fluid flow direction.

44. A semiquantitative method for determining the presence of an analyte in a sample fluid, comprising

exposing a portion of a test strip apparatus to said sample fluid to be tested, wherein said test strip apparatus comprises:

a conjugate pad comprising a diffusible labelled first binding partner that binds with said analyte;

a metering component;

a first area comprising an immobilized second binding partner that competes with said analyte to bind with said diffusible labelled first binding partner; and

a second area comprising an immobilized third binding partner that binds with said diffusible labelled first binding partner,

located in sequence in a capillary fluid flow direction in said test strip apparatus;

allowing said analyte in said sample fluid to migrate up the test strip apparatus by capillary action; and

reading a result of said test strip by comparing relative label intensities in said first area and said second area,

wherein a greater relative label intensity in said second area as compared to said first area indicates the presence of said analyte in said sample fluid in an amount greater than a value determined by the amounts of said first and second binding partners and said metering component and a lesser relative label intensity in said second area as compared to said first area indicates the presence of said analyte in said sample fluid in an amount less than a value determined by the amounts of said first and second binding partners and said metering component.

45. The method of claim 44, wherein said immobilized second binding partner is selected from the group consisting of immobilized analyte and immobilized analyte analog.

46. The method of claim 44, wherein said metering component is contained in said conjugate pad.

47. The method of claim 44, wherein said metering component is contained in said sample pad.

48. The method of claim 44, wherein said metering component is contained in a metering pad located prior to said first area in said capillary fluid flow direction.