Test device for detecting analytes in biological samples

• US 5,656,503 A
• Filed: 09/15/1994
• Issued: 08/12/1997
• Est. Priority Date: 04/27/1987
• Status: Expired due to Term

• Current Assignee: Alere Switzerland GmbH (Abbott Laboratories Incorporated)
• Original Assignee: Unilever Patent Holdings BV (Unilever PLC)
• Inventors: May, Keith, Prior, Michael Evans, Richards, Ian
• Primary Examiner(s): Spiegel, Carol A.
• Application Number:
  US08/305,967
• Time in Patent Office: 1,062 Days
• Field of Search: 422/56-58, 422/60, 427/2, 427/2.11, 435/7.1, 435/7.92-7.95, 435/970, 436/501, 436/514-515, 436/518, 436/523, 436/524, 436/530, 436/541, 436/810, 436/814, 436/525
• US Class Current: 436/514
• CPC Class Codes: G01N 2800/368 Pregnancy complicated by disease or abnormalities of pregnancy, e.g. preeclampsia, preterm labour
• CPC Class Codes: G01N 30/90 Plate chromatography, e.g. thin layer or paper chromatography
• CPC Class Codes: G01N 33/54386 Analytical elements
• CPC Class Codes: G01N 33/54388 based on lateral flow
• CPC Class Codes: G01N 33/548 Carbohydrates, e.g. dextran
• CPC Class Codes: G01N 33/549 with antigen or antibody entrapped within the carrier
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: G01N 33/58 involving labelled substances G01N33/53 takes precedence
• CPC Class Codes: G01N 33/689 related to pregnancy or the gonads
• CPC Class Codes: G01N 33/76 Human chorionic gonadotropin including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors
• CPC Class Codes: Y10S 435/962 Prevention or removal of interfering materials or reactants or other treatment to enhance results, e.g. determining or preventing nonspecific binding
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 435/971 Capture of complex after antigen-antibody reaction
• CPC Class Codes: Y10S 435/973 Simultaneous determination of more than one analyte
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick
• CPC Class Codes: Y10S 436/814 Pregnancy
• CPC Class Codes: Y10S 436/817 Steroids or hormones
• CPC Class Codes: Y10S 436/818 Human chorionic gonadotropin

Abstract

An analytical test device useful for example in pregnancy testing, comprises a hollow casing (500) constructed of moisture-impervious solid material, such as plastics materials, containing a dry porous carrier (510) which communicates indirectly with the exterior of the casing via a bibulous sample receiving member (506) which protrudes from the casing such that a liquid test sample can be applied to the receiving member and permeate therefrom to the porous carrier, the carrier containing in a first zone a labelled specific binding reagent is freely mobile within the porous carrier when in the moist state, wherein the mobility is facilitated by a material comprising a sugar, in an amount effective to reduce interaction between the test strip and the labelled reagent, and in a second zone spatially distinct from the first zone unlabelled specific binding reagent for the same analyte which unlabelled reagent is permanently immobilised on the carrier material and is therefore not mobile in the moist state, the two zones being arranged such that liquid sample applied to the porous carrier can permeate via the first zone into the second zone, and the device incorporating means, such as an aperture (508) in the casing, enabling the extent (if any) to which the labelled reagent becomes bound in the second zone to be observed. Preferably the device includes a removable cap for the protruding bibulous member.

Claims

1. An analytical test device for detecting an analyte suspected of being present in a liquid biological sample selected from the group consisting of urine and serum, said device comprising:

a) a hollow casing having a liquid biological sample application aperture and a test result observation aperture;

b) a bibulous liquid biological sample receiving member within said hollow casing to receive said liquid biological sample applied to said sample application aperture;

c) a test strip comprising a dry porous carrier within said casing and extending from said bibulous liquid biological sample receiving member to and beyond said test result observation aperture, said dry porous carrier having a test result zone observable through said observation aperture;

at least one of said bibulous liquid biological sample receiving member and said test strip containing upstream from said test result zone a labelled reagent capable of specifically binding with said analyte to form a first complex of said labelled reagent and said analyte;

said label being a particulate direct label wherein said labelled reagent is dry on said test strip prior to use and is released into mobile form by said liquid biological sample,

wherein mobility of said labelled reagent within said test strip is facilitated by at least one of

     1) coating at least a portion of said test strip upstream from said test result zone with, or

     2) drying said labelled reagent onto a portion of said test strip upstream from said test zone in the presence of, a material comprising sugar, in an amount effective to reduce interaction between said test strip and said labelled reagent;

said dry porous carrier containing in said test result zone a means for binding said first complex, said means for binding comprising specific binding means and being immobilized in said test result zone;

migration of said liquid biological sample from said bibulous sample receiving member into and through said dry porous carrier conveying by capillarity said first complex to said test result zone of said dry porous carrier whereat said binding means binds said first complex thereby to form a second complex;

said second complex being observable through said test result observation aperture, thereby to indicate the present of said analyte in said liquid biological sample.

2. The test device according to claim 1, wherein said particulate direct label is a sol.

3. The test device according to claim 1, wherein said sol is a dye sol or a metallic sol.

4. The device according to claim 1, wherein said labelled reagent comprises a label selected from the group consisting of dye sols, metallic sols and colored latex particles of maximum diameter not greater than 0.5 microns.

5. The device according to claim 1, wherein said labelled reagent comprises an anti-hCG antibody and said liquid biological sample is urine.

6. The device according to claim 1, wherein said labelled reagent comprises an anti-LH antibody and said liquid biological sample is urine.

7. The analytical test device according to claim 1, wherein said labelled reagent is contained in a first zone of said dry porous carrier upstream from said test result zone, and said means for binding is an unlabelled reagent which is immobilized in said test result zone, said test result zone being spaced from said first zone, said first zone and said test result zone being arranged such that said liquid biological sample applied to said sample aperture is conveyed by capillarity through said first zone into said test result zone.

8. The test device according to claim 1, wherein said hollow casing is molded from plastics material.

9. The test device according to claim 1, wherein said dry porous carrier comprises a strip or sheet of porous material.

10. The test device according to claim 1, wherein said dry porous carrier comprises a strip or sheet of porous material backed with a layer of transparent moisture-impervious material, said transparent layer being in contact with the inside of said hollow casing adjacent said test result observation aperture to inhibit ingress of moisture or said liquid biological sample.

11. The test device according to claim 10, wherein said layer of transparent moisture-impervious material is transparent plastics material.

12. The test device according to claim 1, wherein said dry porous carrier material is nitrocellulose.

13. The test device according to claim 12, wherein said nitrocellulose has a pore size of at least about one micron.

14. The test device according to claim 13, wherein said pore size is about greater than 5 microns.

15. The test device according to claim 14, wherein said pore size is about 8-12 microns.

16. The test device according to claim 1, further comprising a control zone downstream from said test result zone in said dry porous carrier, which control zone comprises means to indicate that said liquid biological sample has been conveyed by capillarity beyond said test result zone, said control zone being observable from outside said hollow casing.

17. The test device according to claim 1, further comprising an absorbent sink at a distal end of said dry porous carrier, said sink having sufficient absorptive capacity to absorb any unbound labelled reagent washed out of said test result zone.

18. The test device according to claim 1, wherein said labelled reagent has been applied to said dry porous carrier as a surface layer.

19. The test device according to claim 18, wherein said dry porous carrier has been pre-treated with a material comprising sugar in the region to which said labelled reagent is applied.

20. The test device according to claim 1, wherein said immobilized reagent in said test result zone is impregnated throughout of said dry porous carrier in said test result zone.

21. The device according to claim 1, wherein said binding means binds directly with said first complex.

22. The device according to claim 1, in combination with a cap for being selectively received over a longitudinal end of said hollow casing to selectively cover said sample application aperture.

23. The device according to claim 1, wherein said bibulous liquid biological sample receiving member protrudes from said hollow casing through said sample application aperture.

24. The device according to claim 1, wherein said dry porous carrier contains said labelled reagent.

25. The device according to claim 1, wherein said bibulous sample receiving member contains said labeled reagent.

26. The test device according to claim 1, wherein said material comprising sugar further comprises a protein.

27. The test device according to claim 26, wherein said protein is bovine serum albumin.

28. The test device according to claim 1, wherein said sugar is selected from the group consisting of sucrose, lactose and dextran.

29. The test device according to claim 1, wherein mobility of said labelled reagent within said test strip is further facilitated by blocking excess binding sites within said test strip with polyvinyl alcohol.

30. The test device according to claim 1, wherein mobility of said labelled reagent within said test strip is further facilitated by blocking excess binding sites within said test strip with a protein.

31. The test device according to claim 30, wherein said protein is selected from the group consisting of bovine serum albumin and milk protein.

32. The test device according to claim 1, wherein said bibulous liquid biological sample receiving member has unidirectional porosity parallel to its longitudinal axis.

33. An analytical method for detecting an analyte suspected of being present in a liquid biological sample selected from the group consisting of urine and serum, which comprises utilizing an analytical test device comprising:

providing an analytical test device having;

a) a hollow casing having a liquid biological sample application aperture and a test result observation aperture;

b) a bibulous liquid biological sample receiving member within said hollow casing to receive said liquid biological sample applied to said sample application aperture;

c) a test strip comprising a dry porous carrier within said hollow casing and extending from said liquid biological sample receiving member to and beyond said test result observation aperture, said dry porous carrier having a test result zone observable through said observation aperture;

said test strip containing in a first zone upstream from said test result zone a labelled reagent capable of specifically binding with said analyte to form a first complex of said labelled reagent and said analyte, said label being a particulate direct label wherein said labelled reagent is dry on said test strip prior to use and is released into mobile form by said liquid biological sample,

wherein mobility of said labelled reagent within said test strip is facilitated by at least one of

     1) coating at least a portion of said test strip upstream from said test result zone with, or

     2) drying said labelled reagent onto a portion of said test strip upstream from said test zone in the presence of, a material comprising sugar, in an amount effective to reduce interaction between said test strip and said labelled reagent;

said carrier containing in said test result zone a means for binding said first complex, said means for binding comprising specific binding means and being immobilized in said test result zone;

migration of said applied liquid biological sample from said bibulous sample receiving member into and through said dry porous carrier conveying by capillarity said first complex to said test result zone of said dry porous carrier whereat said binding means binds said first complex thereby to form a second complex;

said second complex being observable through said test result observation aperture, thereby to indicate the presence of said analyte in said liquid biological sample;

applying said liquid biological sample to said bibulous receiving member;

permitting said liquid biological sample to convey by capillary action through said dry porous carrier via said first zone into said test result zone with said labelled reagent and

detecting the presence of said analyte in said liquid biological sample by visually observing the presence of any labelled reagent bound in said test result zone.

34. The method according to claim 33, wherein said material comprising sugar further comprises a protein.

35. The method according to claim 34, wherein said protein is bovine serum albumin.

36. The method according to claim 33, wherein said sugar is selected from the group consisting of sucrose, lactose and dextran.

37. The method according to claim 33, wherein mobility of said labelled reagent within said test strip is further facilitated by blocking excess binding sites within said test strip with polyvinyl alcohol.

38. The method according to claim 33, wherein mobility of said labelled reagent within said test strip is further facilitated by blocking excess binding sites within said test strip with a protein.

39. The method according to claim 38, wherein said protein is selected from the group consisting of bovine serum albumin and milk protein.

40. The method according to claim 33, wherein said bibulous liquid biological sample receiving member has unidirectional porosity parallel to its longitudinal axis.

41. The method according to claim 40, wherein the bibulous liquid biological sample receiving member protrudes from said hollow casing through said sample application aperture.

42. An analytical test device for detecting an analyte suspected of being present in a liquid biological sample selected from the group consisting of urine and serum, said device comprising:

a) a hollow casing having a liquid biological sample application aperture and a test result observation aperture;

b) a bibulous liquid biological sample receiving member within said hollow casing to receive said liquid biological sample applied to said sample application aperture;

c) a test strip comprising a dry porous carrier within said hollow casing and extending from said bibulous liquid biological sample receiving member to and beyond said test result observation aperture, said dry porous carrier having a test result zone observable through said observation aperture;

at least one of said bibulous liquid biological sample receiving member and said test strip containing upstream from said test result zone a labelled reagent capable of specifically binding with said analyte to form a first complex of said labelled reagent and analyte;

said label being a particulate direct label which is dry prior to use of the device and is released into mobile form by said applied liquid biological sample, wherein mobility of said labelled reagent within said test strip is facilitated by at least one of

     1) coating at least a portion of said test strip upstream from said test result zone with, or

     2) drying said labelled reagent onto a portion of said test strip upstream from said test zone in the presence of, a material comprising sugar, in an amount effective to reduce interaction between said test strip and said labelled reagent;

said carrier containing in said test result zone a means for binding said first complex, said means for binding comprising specific binding means and being immobilized in said test result zone;

(d) a control zone downstream from said test result zone in said dry porous carrier for binding labelled reagent to indicate that said applied liquid biological sample has been conveyed by capillarity beyond said test result zone, said hollow casing having a control zone observation window defined therein, whereby said control zone is observable from outside said hollow casing;

(e) an absorbent sink at a distal end of said carrier, said sink having sufficient absorptive capacity to allow any unbound labelled reagent to wash out of said test result zone;

migration of said applied liquid biological sample from said bibulous sample receiving member into and through said dry porous carrier conveying by capillarity said first complex to said test result zone of said dry porous carrier whereat said binding means binds said first complex thereby to form a second complex;

said second complex being observable through said test result observation aperture, thereby to indicate the presence of said analyte in said applied liquid biological sample.

43. The test device according to claim 42, wherein said material comprising sugar further comprises a protein.

44. The test device according to claim 43 wherein said protein is bovine serum albumin.

45. The test device according to claim 42, wherein said sugar is selected from the group consisting of sucrose, lactose and dextran.

46. The test device according to claim 42, wherein mobility of said labelled reagent within said test strip is further facilitated by blocking excess binding sites within said test strip with polyvinyl alcohol.

47. The test device according to claim 42, wherein mobility of said labelled reagent within said test strip is further facilitated by blocking excess binding sites within said test strip with a protein.

48. The test device according to claim 47, wherein said protein is selected from the group consisting of bovine serum albumin and milk protein.

49. The test device according to claim 42, wherein said bibulous liquid biological sample receiving member has unidirectional porosity parallel to its longitudinal axis.

50. The test device according to claim 49, wherein the bibulous liquid biological sample receiving member protrudes from said hollow casing through said sample application aperture.

51. An analytical test device for detecting an analyte suspected of being present in a liquid biological sample, said device comprising:

a) a hollow casing having a liquid biological sample application aperture and a test result observation aperture;

b) a test strip comprising a dry porous carrier contained within said hollow casing, said carrier communicating directly or indirectly with the exterior of said hollow casing through said liquid biological sample application aperture to receive applied said liquid biological sample, said carrier having a test result zone observable through said test result observation aperture, said dry porous carrier comprising a strip or sheet of porous material backed with a layer of transparent moisture-impervious material, said transparent layer being in contact with the inside of said hollow casing adjacent said test result observation aperture to inhibit ingress of moisture or liquid biological sample, said test strip, in the dry unused state, containing upstream from said test result zone a labelled reagent capable of specifically binding with said analyte to form a first complex of said labelled reagent and said analyte, said label being a particulate direct label which is dry prior to the use of the device and is released into mobile form by said liquid biological sample,

wherein mobility of said labelled reagent within said test strip is facilitated by at least one of

     1) coating at least a portion of said test strip upstream from said test result zone with, or

     2) drying said labelled reagent onto a portion of said test strip upstream from said test zone in the presence of, a material comprising sugar, in an amount effective to reduce interaction between said test strip and said labelled reagent;

said carrier containing in said test result zone a means for binding said first complex, said means for binding comprising specific binding means and being immobilized in said test result zone;

migration of said applied liquid biological sample through said dry porous carrier conveying by capillarity said first complex to said test result zone of said dry porous carrier whereat said binding means binds said first complex thereby to form a second complex;

said second complex being observable through said test result observation aperture, thereby to indicate the presence of said analyte in said liquid biological sample.

52. The test device according to claim 51, wherein said backing material is transparent plastics material.

53. The test device according to claim 51, wherein said dry porous carrier material is nitrocellulose.

54. The test device according to claim 53, wherein said nitrocellulose has a pore size of at least one micron.

55. The test device according to claim 51, wherein said labelled reagent has been applied to said dry porous carrier as a surface layer.

56. The test device according to claim 55, wherein said dry porous carrier has been pre-treated with said material comprising sugar in the region to which said labelled reagent is applied.

57. The test device according to claim 51, wherein said analyte is hCG.

58. The test device according to claim 51, wherein said analyte is LH.

59. The test device according to claim 51, wherein said material further comprises a protein.

60. The test device according to claim 59 wherein said protein is bovine serum albumin.

61. The test device according to claim 51, wherein said sugar is selected from the group consisting of sucrose, lactose and dextran.

62. The test device according to claim 51, wherein mobility of said labelled reagent within said test strip is further facilitated by blocking excess binding sites within said test strip with polyvinyl alcohol.

63. The test device according to claim 51, wherein mobility of said labelled reagent within said test strip is further facilitated by blocking excess binding sites within said test strip with a protein.

64. The test device according to claim 63, wherein said protein is selected from the group consisting of bovine serum albumin and milk protein.