Opposable-element assay devices, kits, and methods employing them

• US 5,998,220 A
• Filed: 02/10/1994
• Issued: 12/07/1999
• Est. Priority Date: 05/29/1991
• Status: Expired due to Term

• Current Assignee: Beckman Coulter Incorporated (Danaher Corporation)
• Original Assignee: Beckman Coulter Incorporated (Danaher Corporation)
• Inventors: Chandler, Howard M.
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US08/194,793
• Time in Patent Office: 2,126 Days
• Field of Search: 422/55, 422/56, 422/58, 422/61, 435/7.92, 435/7.93, 435/7.94, 435/287.1, 435/287.2, 435/287.7, 435/287.8, 435/287.9, 435/288.5, 435/805, 435/810, 435/970, 435/973, 435/975, 436/514, 436/518, 436/530, 436/164, 436/169, 436/807, 436/808, 436/810
• US Class Current: 436/514
• CPC Class Codes: B01D 15/3804 Affinity chromatography
• CPC Class Codes: B01L 2300/0825 Test strips
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 3/5023 with a sample being transported to, and subsequently stored in an absorbent for analysis
• CPC Class Codes: B01L 3/5055 Hinged, e.g. opposable surfaces
• CPC Class Codes: G01N 30/02 Column chromatography
• CPC Class Codes: G01N 30/30 of temperature
• CPC Class Codes: G01N 30/90 Plate chromatography, e.g. thin layer or paper chromatography
• CPC Class Codes: G01N 33/5302 Apparatus specially adapted for immunological test procedures
• 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 435/805 Test papers
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 435/969 Multiple layering of reactants
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 435/973 Simultaneous determination of more than one analyte
• CPC Class Codes: Y10S 435/975 Kit
• 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 436/809 Multifield plates or multicontainer arrays
• CPC Class Codes: Y10S 436/81 : Tube, bottle, or dipstick

Abstract

A chromatographic assay device for use with immunoassays allows rapid and convenient assays of analytes of biological interest, and permits extractions to be carried out in situ, avoiding the use of separate extraction vessels. The device has a wide dynamic range and avoids interference from particulates or colored components. In one form, the device comprises: (1) a first opposable component comprising a sample preparation means adapted to receive a sample to be assayed; and (2) a second opposable component comprising a chromatographic medium. The first and second opposable components can be brought into opposition so as to cause the sample preparation means to apply the sample to be tested to the chromatographic medium. Preferably, the analyte is detected with a visually detectable label. Other variations of the device vary the arrangement of components to provide optimal chromatography for a variety of analytes, as well as to permit bi-directional chromatography. The devices can be incorporated in test kits, and assay methods using the devices are also disclosed.

Claims

1. A chromatographic assay device for detection and/or determination of an analyte comprising:

(a) a first opposable component including a sample preparation means for receiving a sample to be assayed; and

(b) a second opposable component including a chromatographic medium including thereon a detection zone containing an immobilized specific binding partner for binding the analyte to be detected and/or determined;

wherein the first and second opposable components can be brought into operable contact so as to cause the sample preparation means to apply the sample to be tested to the chromatographic medium.

2. The chromatographic assay device of claim 1 wherein the sample preparation means contains at least one reagent for treatment of the sample before the sample is applied to the chromatographic medium.

3. The chromatographic assay device of claim 2 wherein the reagent for treatment of the sample is an extraction reagent to extract analyte from the sample.

4. The chromatographic assay device of claim 1 wherein the chromatographic medium has first and second ends and the device further comprises a conducting means in operable contact with the first end of the chromatographic medium.

5. The chromatographic assay device of claim 4 further comprising an absorbing means in operable contact with the second end of the chromatographic medium.

6. The chromatographic assay device of claim 1 wherein the chromatographic medium has first and second ends and the detection zone is smaller than the chromatographic medium.

7. The chromatographic assay device of claim 6 wherein the chromatographic medium further includes a control zone smaller than the chromatographic medium and separate from the detection zone so that the binding of a labeled specific binding partner can be detected at both the control zone and the detection zone, the control zone being located closer to the second end of the chromatographic medium than the detection zone.

8. The chromatographic assay device of claim 7 wherein the control zone contains analyte immobilized thereto.

9. The chromatographic assay device of claim 1 wherein the chromatographic medium further includes a control zone substantially smaller than the chromatographic medium.

10. A test kit for the detection and/or determination of an analyte comprising, separately packaged:

(a) the chromatographic assay device of claim 1; and

(b) a specific binding partner for the analyte labeled with a detectable label, the specific binding partner to be used with the chromatographic assay device.

11. The test kit of claim 10 wherein the detectable label is a visually detectable label.

12. A chromatographic assay device for detection and/or determination of an analyte comprising:

(a) a first opposable component including a sample preparation means for receiving a sample to be assayed; and

(b) a second opposable component including a chromatographic medium including thereon a detection zone of an immobilized specific binding partner for binding the analyte to be detected and/or determined;

wherein the first and second opposable components can be brought into operable contact so as to cause the sample preparation means to apply the sample to be tested to the chromatographic medium and wherein the first and second opposable components each further include engaging means for securing the first and second opposable components in opposition.

13. A chromatographic assay device for detecting and/or determination of an analyte comprising:

(a) a first opposable component including a sample preparation means for receiving a sample to be assayed; and

(b) a second opposable component including a chromatographic medium including thereon a detection zone containing an immobilized specific binding partner for binding the analyte to be detected and/or determined;

wherein the first and second opposable components can be brought into operable contact so as to cause the sample preparation means to apply the sample to be tested to the chromatographic medium and wherein the first and second opposable components are joined by a hinge.

14. A chromatographic assay device for detection and/or determination of an analyte comprising:

(a) a first opposable component including;

(i) a chromatographic medium having first and second ends and including thereon a detection zone containing an immobilized specific binding partner for binding the analyte to be detected and/or determined; and

(ii) a first application means at the first end of the chromatographic medium; and

(b) a second opposable component including;

(i) a second application means for receiving a second liquid; and

(ii) an absorbing means separated from the second application means;

wherein addition of a first liquid to the first application means causes the first liquid to be applied to the first end of the chromatographic medium; and

wherein bringing the first and second opposable components into operable contact;

(i) causes the second application means to come into operable contact with the second end of the chromatographic medium so as to apply a second liquid to the second end of the chromatographic medium; and

(ii) causes the absorbing means to come into operable contact with the first application means so as to withdraw fluid from the chromatographic medium via the first application means.

15. The chromatographic assay device of claim 14 wherein the first application means comprises a sample preparation means containing at least one reagent for treatment of the sample before the sample is applied to the chromatographic medium.

16. The chromatographic assay device of claim 14 further comprising a conductive medium in operable contact with the second end of the chromatographic medium.

17. The chromatographic assay device of claim 14 wherein the first application means is impregnated with an inert dye.

18. A test kit for the detection and/or determination of an analyte comprising, separately packaged:

(a) the chromatographic assay device of claim 14; and

(b) a specific binding partner for the analyte labeled with a detectable label, the specific binding partner to be used with the chromatographic assay device.

19. The test kit of claim 18 wherein the detectable label is a visually detectable label.

20. A method for detecting and/or determining an analyte in a sample comprising the steps of:

(a) applying the sample as the first liquid to the first application means of the chromatographic assay device of claim 14;

(b) allowing the sample to move through at least a portion of the chromatographic medium, the portion including the detection zone;

(c) applying a detection reagent as the second liquid to the second application means, the detection reagent comprising at least one component capable of binding specifically to any analyte present in the sample;

(d) bringing the first and second opposable components into operable contact, thereby causing the second application means to come into operable contact with the second end of the chromatographic medium so as to apply the second liquid to the second end of the chromatographic medium and causing the absorbing means to come into operable contact with the first application means so as to cause the detection reagent to move through at least a portion of the chromatographic medium including the detection zone, so that the detection reagent gives a detectable indication of the presence and/or quantity of the analyte; and

(e) observing and/or measuring the detection reagent at the detection zone in order to detect and/or determine the analyte.

21. The method of claim 20 wherein the detection reagent comprises a specific binding partner for the analyte labelled with a detectable label.

22. The method of claim 21 wherein the detectable label is a visually detectable label and the step of observing and/or measuring the detection reagent comprises visually observing the detection reagent.

23. A chromatographic assay device for detection and/or determination of an analyte comprising:

(a) a first opposable component including;

(i) a sample preparation means for receiving a sample to be assayed; and

(ii) a chromatographic medium that is not in operable contact with the sample preparation means, the chromatographic medium including thereon a detection zone containing an immobilized specific binding partner for binding the analyte to be detected and/or determined; and

(b) a second opposable component including a fluid-conducting connecting member;

wherein, when the first and second opposable components are brought into operable contact, the connecting member is brought into operable contact with both the sample preparation means and the chromatographic medium to result in the application of the sample to the chromatographic medium.

24. The chromatographic assay device of claim 23 wherein the sample preparation means contains at least one reagent for treatment of the sample before the sample is applied to the chromatographic medium.

25. The chromatographic assay device of claim 23 wherein the chromatographic medium has first and second ends and the device further comprises a conducting means in operable contact with the second end of the chromatographic medium.

26. The chromatographic assay device of claim 23 the detection zone is smaller than the chromatographic medium.

27. The chromatographic assay device of claim 26 wherein the chromatographic medium further includes a control zone smaller than the chromatographic medium and separate from the detection zone so that a labeled specific binding partner bound to the detection zone and a labeled specific binding partner bound to the control zone can be detected simultaneously, the control zone being located closer to the second end of the chromatographic medium than the detection zone.

28. A test kit for the detection and/or determination of an analyte comprising, separately packaged:

(a) the chromatographic assay device of claim 23; and

(b) a specific binding partner for the analyte labeled with a detectable label, the specific binding partner to be used with the chromatographic assay device.

29. The test kit of claim 28 wherein the detectable label is a visually detectable label.

30. A chromatographic assay device for detection and/or determination of an analyte comprising:

(a) a first opposable component including;

(i) a sample preparation means for receiving a sample to be assayed; and

(ii) a chromatographic medium that is not in operable contact with the sample preparation means, the chromatographic medium including a detection zone containing an immobilized specific binding partner for binding the analyte that is to be detected and/or determined; and

(b) a second opposable component including a fluid-conducting connecting member;

wherein, when the first and second opposable components are brought into operable contact, the connecting member is brought into operable contact with both the sample preparation means and the chromatographic medium so as to result in the application of the sample to the chromatographic medium and wherein the first and second opposable components each further comprise engaging means for securing the first and second opposable components in operable contact.

31. A chromatographic assay device for detection and/or determination of an analyte comprising:

(a) a first opposable component including;

(i) a chromatographic medium having first and second ends and having thereon a detection zone containing an immobilized specific binding partner for binding the analyte to be detected and/or determined;

(ii) a first application means in operable contact with the first end of the chromatographic medium; and

(iii) a first absorbing means of finite capacity in operable contact with the second end of the chromatographic medium; and

(b) a second opposable component including;

(i) a second application means; and

(ii) a second absorbing means;

wherein the first and second opposable components can be brought into operable contact so that the second application means is placed in operable contact with the first absorbing means and the second absorbing means is placed in operable contact with the first application means.

32. The chromatographic assay device of claim 31 wherein the first application means comprises a sample application pad and the second application means comprises a detector application pad, to which detecting reagent can be applied, whereby a sample applied to the sample application pad first flows through the chromatographic medium from the first end to the second end, and, when the first and second opposable components are brought into opposition, a detecting reagent applied to the detector application pad flows through the chromatographic medium from the second end to the first end.

33. The chromatographic assay device of claim 32 wherein the sample application pad contains at least one reagent for treatment of a sample before the sample is applied to the chromatographic medium.

34. The chromatographic assay device of claim 31 the detection zone is smaller in area than the chromatographic medium.

35. A test kit for the detection and/or determination of an analyte comprising, separately packaged:

(a) the chromatographic assay device of claim 31; and

(b) a specific binding partner for the analyte labeled with a detectable label, to be applied to the second application means of the chromatographic assay device.

36. A method for detecting and/or determining an analyte in a sample comprising the steps of:

(a) applying the sample to the first application means of the chromatographic assay device of claim 31 so that the sample flows through at least a portion of the chromatographic medium from the first end toward the second end, the portion including the detection zone;

(b) applying a detection reagent to the second application means of the chromatographic assay device;

(c) bringing the first and second opposable components into operable contact so that the detection reagent is applied to the chromatographic medium;

(d) allowing the detection reagent to move through at least a portion of the chromatographic medium, the portion including the detection zone, from the second end toward the first end so that the detection reagent gives a detectable indication of the presence and/or quantity of the analyte; and

(e) observing and/or measuring the detection reagent at the detection zone in order to detect and/or determine the analyte.

37. The method of claim 36 wherein the detection reagent comprises a visually detectable label and the step of observing and/or measuring the detection reagent comprises visually observing the detection reagent.

38. A chromatographic assay device for detection and/or determination of an analyte comprising:

(a) a first opposable component including;

(i) a chromatographic medium having first and second ends and including thereon a detection zone containing an immobilized specific binding partner for binding the analyte to be detected and/or determined;

(ii) a first application means in operable contact with the first end of the chromatographic medium and located in a recess of the first opposable component; and

(iii) a first absorbing means in operable contact with the second end of the chromatographic medium;

(b) a second opposable component including;

(i) a second absorbing means; and

(ii) a second application means separated from the second absorbing means; and

(c) a cover hingedly attached to the first opposable component so that it can be folded over the first and second opposable components when they are opposed;

wherein addition of a first liquid to the first application means causes the first liquid to be applied to the first end of the chromatographic medium; and

wherein bringing the first and second opposable components into operable contact;

(i) causes the second application means to come into operable contact with the first absorbing means so as to apply a second liquid to the second end of the chromatographic medium; and

(ii) causes the second absorbing means to come into operable contact with the first application means and into direct contact with at least a portion of the chromatographic medium to withdraw fluid from the chromatographic medium via the first application means.

39. The chromatographic assay device of claim 38 wherein the first application means further comprises a sample preparation means containing at least one reagent for treatment of the sample before the sample is applied to the chromatographic medium.

40. The chromatographic assay device of claim 38 wherein the second opposable component and the cover each have an aperture cut therein to permit viewing of at least a portion of the chromatographic medium when the first and second opposable components are opposed and the cover is folded over the first and second opposable components.

41. The chromatographic assay device of claim 38 the detection zone is smaller than the chromatographic medium.

42. A test kit for the detection and/or determination of an analyte comprising, separately packaged:

(a) the chromatographic assay device of claim 38; and

(b) a specific binding partner for the analyte labeled with a detectable label, the specific binding partner to be used with the chromatographic assay device.

43. The test kit of claim 42 wherein the detectable label is a visually detectable label.

44. The test kit of claim 42 wherein the specific binding partner for the analyte labelled with the detectable label is located in the second application means in a form that can be resolubilized by the addition of an aqueous liquid to the second application means.

45. A method for detecting and/or determining an analyte in a sample comprising the steps of:

(a) applying the sample as the first aqueous liquid to the first application means of the chromatographic assay device of claim 44;

(b) allowing the sample to move through at least a portion of the chromatographic medium, the portion including the detection zone;

(c) applying a second aqueous liquid to the second application means, thereby resolubilizing the specific binding partner for the analyte labeled with a detectable label, the labeled specific binding partner resolubilized in the second liquid forming a detection reagent;

(d) bringing the first and second opposable components into operable contact, thereby causing the second application means to come into operable contact with the second end of the chromatographic medium, so as to apply the detection reagent to the second end of the chromatographic medium and causing the absorbing means to come into operable contact with the first application means, so as to cause the detection reagent to move through at least a portion of the chromatographic medium including the detection zone, so that the detection reagent gives a detectable indication of the presence and/or quantity of the analyte;

(e) folding the cover hingedly attached to the first opposable component over the first and second opposable components; and

(f) observing and/or measuring the detection reagent at the detection zone in order to detect and/or determine the analyte.

46. The method of claim 45 wherein the detectable label is a visually detectable label and the step of observing and/or measuring the detection reagents comprises visually observing the detection reagent.

47. A method for detecting and/or determining an analyte in a sample comprising the steps of:

(a) applying the sample as the first liquid to the first application means of the chromatographic assay device of claim 38;

(b) allowing the sample to move through at least a portion of the chromatographic medium, the portion including the detection zone;

(c) applying the detection reagent as the second liquid to the second application means, the detection reagent comprising at least one component capable of binding specifically to any analyte present in the sample;

(d) bringing the first and second opposable components into operable contact, thereby causing the second application means to come into operable contact with the first absorbing means so as to apply the second liquid to the second end of the chromatographic medium and causing the second absorbing means to come into contact with the first application means so as to withdraw fluid from the chromatographic medium via the first application means, thereby causing the detection reagent to move through at least a portion of the chromatographic medium including the detection zone, so that the detection reagent gives a detectable indication of the presence and/or quantity of the analyte;

(e) folding the cover over the first and second opposable components; and

(f) observing and/or measuring the detection reagent at the detection zone in order to detect and/or determine the analyte.

48. The method of claim 47 wherein the detectable label is a visually detectable label and the step of observing and/or measuring the detection reagent comprises visually observing the detection reagent.

49. A chromatographic assay device for the detection and/or determination of an analyte comprising:

(a) a first opposable component including;

(i) a chromatographic medium having first and second ends and including a detection zone containing an immobilized specific binding partner for binding the analyte to be detected and/or determined;

(ii) a first conducting means in operable contact with the first end of the chromatographic medium; and

(iii) a second conducting means in operable contact with the second end of the chromatographic medium;

(b) a second opposable component hingedly attached to the first opposable component including;

(i) a first absorbing means; and

(ii) a first application means separated from the first absorbing means;

wherein bringing the first and second opposable components into operable contact causes the first absorbing means to come into operable contact with the second conducting means to withdraw fluid from the chromatographic medium, and causes the first application means to come into operable contact with the first conducting means to apply fluid to the chromatographic medium, so that a first liquid applied to the first application means is drawn through at least a portion of the chromatographic medium including the detection zone; and

(c) a third opposable component hingedly attached to the first opposable component including;

(i) a second absorbing means; and

(ii) a second application means separated from the second absorbing means;

wherein bringing the first and third opposable components into operable contact causes the second absorbing means to come into operable contact with the first conducting means to withdraw fluid from the chromatographic medium and causes the second application means to come into operable contact with the second conducting means to apply fluid to the chromatographic medium so that a second liquid applied to the second application means is drawn through at least a portion of the chromatographic medium overlapping the portion of the chromatographic medium through which the first liquid is drawn, including the detection zone;

the first, second, and third opposable components being in such a configuration that, when the third opposable component is brought into operable contact with the first opposable component, the second opposable component can be folded over the first and third opposable components to form a cover.

50. The chromatographic assay device of claim 49 wherein the first application means comprises a sample application pad containing at least one reagent for the treatment of a sample before the sample is applied to the chromatographic medium.

51. The chromatographic assay device of claim 49 wherein the first application means contains a first specific binding partner for the analyte in a form that can be resolubilized by the application of an aqueous sample to the application means.

52. A method for detecting and/or determining an analyte in a sample comprising the steps of:

(a) applying the sample to the chromatographic assay device of claim 51, thereby resolubilizing the first specific binding partner for the analyte and forming a solution containing a complex of the analyte and the first specific binding partner for the analyte;

(b) bringing the first and second opposable components into operable contact so that the first application means applies the complex of the analyte and the first specific binding partner to the analyte to the first conducting means;

(c) allowing the complex of the analyte and the first specific binding partner to the analyte to move through at least a portion of the chromatographic medium, including the detection zone;

(d) separating the first and second opposable components so that they are no longer in operable contact;

(e) adding a fluid to the second application means located on the third opposable component, the second application means containing a specific binding partner to the first specific binding partner to the analyte, the specific binding partner to the first specific binding partner to the analyte labelled with a detectable label, the specific binding partner contained on the second application means being present in a form that can be resolubilized by the addition of the fluid to the second application means;

(f) bringing the first and third opposable components into operable contact, thereby causing the resolubilized labeled specific binding partner in the second application means to be drawn through at least a portion of the chromatographic medium including the detection zone; and

(g) observing and/or measuring the detection reagent at the detection zone in order to detect and/or determine the analyte.

53. The method of claim 52 wherein the detectable label is a visually detectable label and the step of observing and/or measuring the detectable label comprises visually observing the visually detectable label.

54. The chromatographic assay device of claim 49 wherein the second liquid comprises a specific binding partner to the analyte and the chromatographic medium further comprises a detection zone containing a secondary specific binding partner, the secondary specific binding partner being specific for the first specific binding partner.

55. A test kit for the detection and/or determination of an analyte comprising. separately packaged:

(a) the chromatographic assay device of claim 49; and

(b) a specific binding partner for the first specific binding partner to the analyte labeled with a detectable label, the specific binding partner for the first specific binding partner to the analyte to be used with the chromatographic assay device.

56. The test kit of claim 55 wherein the detectable label is a visually detectable label.

57. A chromatographic assay device for detection and/or determination of at least one analyte comprising:

(a) a first opposable component including a plurality of laterally separated sample preparation means, each for receiving a sample to be assayed; and

(b) a second opposable component including a chromatographic medium for each sample preparation means on the first opposable component, each chromatographic medium including thereon a detection zone containing an immobilized specific binding partner for the analyte, the chromatographic media being laterally separated;

wherein the first and second opposable components can be brought into operable contact so as to cause each sample preparation means to apply each sample to be tested to the corresponding chromatographic medium.

58. The chromatographic assay device of claim 57 wherein each detection zone is smaller than the chromatographic medium.

59. The chromatographic assay device of claim 57 wherein each of the immobilized specific binding partners is specific for the same analyte.

60. The chromatographic assay device of claim 57 wherein at least one sample preparation means includes a collapsible well for receiving a sample-containing device.

61. The chromatographic assay device of claim 60 wherein the first opposable component further includes hingedly foldable wings that fold over the second opposable component when the first opposable component and second opposable component are brought into opposition.

62. A test kit for the detection and/or determination of at least one analyte comprising, separately packaged:

(a) the chromatographic assay device of claim 57; and

(b) a specific binding partner for each analyte to be tested, each specific binding partner labeled with a detectable label, each specific binding partner packaged separately, and each specific binding partner to be used with the chromatographic assay device.

63. The test kit of claim 62 wherein each detectable label is a visually detectable label.

64. A chromatographic assay device for detection and/or determination of an analyte comprising:

(a) a first opposable component including;

(i) a chromatographic medium having first and second ends and including a detection zone containing an immobilized specific binding partner for binding the analyte to be detected and/or determined;

(ii) a conducting means in operable contact with the first end of the chromatographic medium; and

(iii) an absorbing means in operable contact with the second end of the chromatographic medium; and

(b) a second opposable component comprising;

(i) a first application means; and

(ii) a second application means;

the first and second application means being positioned on the second opposable component such that they are not in operable contact when the first and second opposable components are not in operable contact;

wherein bringing the first and second opposable components into operable contact places the conducting means in operable contact with the first application means and places the conducting means in operable contact with the second application means, thereby placing the first and second application means in operable contact with each other.

65. A test kit for the detection and/or determination of an analyte comprising, separately packaged:

(a) the chromatographic assay device of claim 64; and

(b) a specific binding partner for the analyte labeled with a detectable label, to be applied to the second application means of the chromatographic assay device.

66. A chromatographic assay device for the detection and/or determination of an analyte comprising:

(a) a first opposable component including;

(i) a chromatographic medium having first and second ends and including thereon a detection zone containing an immobilized specific binding partner for binding the analyte to be detected and/or determined;

(ii) a conducting means in operable contact with the first end of the chromatographic medium;

(iii) an absorbing means in operable contact with the second end of the chromatographic medium; and

(iv) a detector application pad in direct contact with the conducting means and positioned such that it is in indirect contact with the first end of the chromatographic medium; and

(b) a second opposable component including a sample application pad;

whereby bringing the first and second opposable components into operable contact causes the sample application pad to apply the sample to be tested to the detector application pad and thus the first end of the chromatographic medium through the conducting means.

67. A chromatographic assay device for the detection and/or determination of an analyte comprising:

(a) a first opposable component including;

(i) a chromatographic medium having first and second ends and including thereon a detection zone containing an immobilized specific binding partner for binding the analyte to be detected and/or determined;

(ii) an absorbing means in operable contact with the second end of the chromatographic medium; and

(iii) a detector application pad in operable contact with the first end of the chromatographic medium; and

(b) a second opposable component including a sample application pad;

wherein the first and second opposable components are brought into operable contact, the detector application pad and the sample application pad are in contact except for the region of the detector application pair directly adjacent to the first end of the chromatographic medium, and whereby bringing the first and second opposable components into operable contact causes the sample application pad to apply the sample to be tested to the detector application pad and thus to the first end of the chromatographic medium.

68. A test kit for the detection and/or determination of an analyte comprising, separately packaged:

(a) the chromatographic assay device of claim 67; and

(b) a specific binding partner for the analyte labeled with a detectable label, to be applied to the second application means of the chromatographic assay device.

69. A chromatographic assay device for detection and/or determination of an analyte comprising:

(a) a first opposable component including;

(i) a chromatographic medium having first and second ends and including thereon a detection zone containing an immobilized specific binding partner for binding the analyte to be detected and/or determined;

(ii) a conducting means positioned such that it is not in operable contact with the first end of the chromatographic medium when the first opposable component and second opposable component are not in operable contact; and

(iii) an absorbing means in operable contact with the second end of the chromatographic medium; and

(b) a second opposable component comprising;

(i) a first application means for application of a first liquid; and

(ii) a second application means for application of a second liquid;

the first and second application means being positioned on the second opposable component such that they are not in operable contact when the first and second opposable components are not in operable contact;

wherein bringing the first and second opposable components into operable contact places the conducting means in operable contact with the first application means, places the conducting means in operable contact with the second application means, and places the second application means in operable contact with the first end of the chromatographic medium, thereby placing-the first and second application means in operable contact with each other to apply the first and second liquids to the chromatographic medium.

70. A test kit for the detection and/or determination of an analyte comprising, separately packaged:

(a) the chromatographic assay device of claim 69; and

(b) a specific binding partner for the analyte labeled with a detectable label, to be applied to the second application means of the chromatogranhic assay device.

71. A chromatographic assay device for detection and/or determination of an analyte comprising:

(a) a first opposable component including;

(i) a sample preparation means for receiving a sample to be assayed; and

(ii) a chromatographic medium that is not in operable contact with the sample preparation means and including a detection zone containing an immobilized specific binding partner for binding the analyte to be detected and/or determined; and

(b) a second opposable component including a fluid-conducting connecting member;

wherein, when the first and second opposable components are brought into operable contact, the connecting member is brought in operable contact with both the sample preparation means and the chromatographic medium so as to result in the application of the sample to the chromatographic medium and wherein the first and second opposable components are joined by a hinge.