Assays

• US 5,602,040 A
• Filed: 05/12/1994
• Issued: 02/11/1997
• Est. Priority Date: 04/27/1987
• Status: Expired due to Term

• Current Assignee: Inverness Medical Switzerland GmbH (Abbott Laboratories Incorporated)
• Original Assignee: Unilever Patent Holdings BV (Unilever PLC)
• Inventors: Prior, Michael E., May, Keith, Richards, Ian
• Primary Examiner(s): Spiegel, Carol A.
• Application Number:
  US08/241,675
• Time in Patent Office: 1,006 Days
• Field of Search: 422/56-58, 422/60, 436/501, 436/530, 436/514, 436/810, 436/814, 436/515, 436/518, 436/523, 436/524, 436/541, 436/817, 436/818, 436/906, 435/7.92-7.95, 435/970, 435/810, 435/962, 435/975, 427/2, 427/2.11, 427/2.13
• 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 33/54366 Apparatus specially adapted for solid-phase testing
• 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 435/805 Test papers
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• 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 435/969 Multiple layering of reactants
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 435/975 Kit
• 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
• CPC Class Codes: Y10S 436/906 : Fertility tests

Abstract

An analytical test device useful for example in pregnancy testing, includes 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 sugar, in a amount effective to reduce interaction between the test strip and the labelled reagent, and in a second zone spatially distinct from the first zone unlabeled specific binding reagent for the same analyte which unlabelled reagent is permanently immobilized 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 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, said device comprising:

a) a hollow casing having a first end, a second end, a longitudinal axis, and a liquid biological sample application aperture adjacent to said first end;

b) a displaceable shroud disposed on said first end for shielding said liquid biological sample application aperture;

c) a test strip comprising a dry porous carrier disposed within said hollow casing, said dry porous 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 dry porous carrier having a detection zone;

said hollow casing including means permitting observation of said detection zone from outside said casing;

said test strip, in the dry unused state, containing upstream from said detection zone a labelled reagent capable of specifically binding with said analyte to form a first complex of said labelled reagent and said analyte, said labelled reagent being labelled with 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 detection zone with, or

     2) drying said labelled reagent onto a portion of said test strip upstream of said detection 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 detection zone a means for binding said first complex, said means for binding comprising specific binding means and being immobilized in said detection zone;

migration of said liquid biological sample through said dry porous carrier conveying by capillarity said first complex to said detection 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 via said detection zone observation means, thereby to indicate the presence of said analyte in said liquid biological sample.

2. The test device according to claim 1, wherein said particulate direct label is selected from the group consisting of a dye sol, a metallic sol, and colored latex particles having a maximum diameter of less than or equal to about 0.5 micron.

3. The test device according to claim 1, wherein said means permitting observation comprises at least one aperture through which the second complex may be observed.

4. The test device according to claim 3, 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 at least one aperture to inhibit ingress of moisture or said liquid biological sample.

5. The test device according to claim 1, wherein said hollow casing and said shroud are molded from a plastic material.

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

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

8. The test device according to claim 7, wherein said nitrocellulose has a pore size of at least about one micron.

9. The test device according to claim 7, wherein said pore size is greater than about 5 microns.

10. The test device according to claim 7, wherein said pore size is about 8-12 microns.

11. The test device according to claim 1, incorporating a control zone downstream from said detection 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 detection zone, said hollow casing further including means permitting observation of said control zone from outside said hollow casing.

12. 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 allow any labelled reagent not bound with said analyte to wash out of said detection zone.

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

14. The test device according to claim 13, wherein said dry porous carrier has been pre-treated with said material comprising sugar in the upstream portion to which said labelled reagent is applied.

15. The test device according to claim 1, wherein said means for binding in said detection zone is impregnated throughout the thickness of said dry porous carrier in said detection zone.

16. The test device according to claim 1, wherein said analyte is hCG.

17. The test device according to claim 1, wherein said analyte is LH.

18. The test device according to claim 1, wherein

(a) said dry porous carrier

(i) is nitrocellulose and

(ii) communicates indirectly with the exterior of said hollow casing via a bibulous urine receiving member which protrudes from said hollow casing and which acts as a reservoir from which urine is released;

(b) contains said labelled reagent in a first zone upstream from said detection zone, wherein said labelled reagent

(i) comprises an anti-LH antibody conjugated to a colored particulate label and

(ii) is freely mobile within said dry porous carrier when wetted with said urine;

(c) said detection zone is spatially distinct from said first zone;

(d) said means for binding said first complex is an unlabelled anti-LH antibody which is permanently immobilized in said detection zone, wherein said unlabelled anti-LH antibody specifically binds a different LH epitope than the anti-LH antibody of said labelled reagent; and

(e) said first zone and said detection zone being arranged such that said urine applied to said protruding bibulous urine receiving member, and consequently thereby to said dry porous carrier, is conveyed by capillarity via said first zone into said detection zone, said hollow casing having at least one aperture through which said second complex can be observed, whereby said test device is a fertile period prediction device.

19. The test device according to claim 1, wherein

(a) said dry porous carrier

(i) is nitrocellulose and

(ii) communicates indirectly with the exterior of said hollow casing via a bibulous urine receiving member which protrudes from said hollow casing and which acts as a reservoir from which urine is released;

(b) contains said labelled reagent in a first zone upstream from said detection zone, wherein said labelled reagent

(i) comprises an anti-hCG antibody conjugated to a colored particulate label and

(ii) is freely mobile within said dry porous carrier when wetted with said urine;

(c) said detection zone is spatially distinct from said first zone;

(d) said means for binding said first complex is an unlabelled anti-hCG antibody which is permanently immobilized in said detection zone, wherein said unlabelled anti-hCG antibody specifically binds a different hCG epitope than the anti-hCG antibody of said labelled reagent; and

,

(e) said first zone and said detection zone being arranged such that said urine applied to said protruding bibulous urine receiving member, and consequently thereby to said dry porous carrier, is conveyed by capillarity via said first zone into said detection zone, said hollow casing having at least one aperture through which said second complex can be observed, whereby said test device is a pregnancy test device.

20. The device according to claim 1, wherein said test strip further comprises a bibulous sample receiving member for initially receiving said liquid biological sample, said bibulous sample receiving member thereafter releasing said liquid biological sample into said dry porous carrier, and wherein said dry porous carrier contains said labelled reagent.

21. The test device according to claim 20, wherein said bibulous sample receiving member protrudes from said housing through said sample aperture, and wherein said shroud surrounds said protruding bibulous member.

22. The test device according to claim 1, which further comprises a control zone downstream from said detection 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 detection zone, said hollow casing further including means permitting observation of said control zone from outside said hollow casing.

23. The test device according to claim 22, wherein said means permitting observation of said control zone comprises a control zone observation aperture.

24. The test device according to claim 1, wherein said means permitting observation of said detection zone comprises a detection zone observation aperture.

25. The test device according to claim 1, wherein said shroud is constructed of opaque or translucent material.

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 20, wherein said bibulous sample receiving member has unidirectional porosity parallel to its longitudinal axis.

33. A kit comprising a packaged pregnancy test device for detecting hCG suspected of being present in a urine sample, said device comprising:

a) a hollow casing having a first end, a second end, a longitudinal axis, and a urine sample application aperture adjacent to said first end;

b) a displaceable shroud disposed on said first end for shielding said urine sample application aperture;

c) a test strip comprising a dry porous carrier disposed within said hollow casing, wherein said dry porous carrier is nitrocellulose and communicates indirectly with the exterior of said hollow casing via a bibulous urine sample receiving member which protrudes from said hollow casing and which acts as a reservoir from which said urine sample is released into said dry porous carrier,

said dry porous carrier having a detection zone;

said hollow casing including means permitting observation of said detection zone from outside said casing;

said dry porous carrier containing in a first zone upstream from said detection zone a labelled reagent comprising a colored particulate direct label conjugated to an anti-hCG antibody, wherein said anti-hCG antibody present in said labelled reagent is capable of specifically binding with said hCG to form a first complex of said labelled reagent and said hCG,

wherein said labelled reagent is dry on said test strip prior to use and is released into mobile form by said urine sample, said labelled reagent being freely mobile within said test strip when said test strip is wetted with said urine sample, said detection zone being spatially distinct from said first zone;

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

     1) coating at least said first zone with, or

     2) drying said labelled reagent onto a portion of said first 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 detection zone a means for specifically binding said first complex, said means for specifically binding said first complex being an unlabelled anti-hCG antibody which is impregnated throughout the thickness of said dry porous carrier in said detection zone and is permanently immobilized in said detection zone,

wherein said anti-hCG antibody of said labelled reagent and said unlabelled anti-hCG antibody specifically bind to different hCG epitopes,

said first zone and said detection zone being arranged such that said urine sample applied to said protruding bibulous urine sample receiving member and consequently to said dry porous carrier can migrate via said first zone into said detection zone,

said migration of said urine sample through said dry porous carrier conveying by capillarity said first complex to said detection zone whereat said means for specifically binding binds said first complex thereby to form a second complex;

said second complex being observable via said means permitting observation in said detection zone, thereby to indicate the presence of said hCG in said urine sample.

34. A kit comprising a packaged fertile period prediction test device for detecting LH suspected of being present in a urine sample, said device comprising:

a) a hollow casing having a first end, a second end, a longitudinal axis, and a urine sample application aperture adjacent to said first end;

b) a displaceable shroud disposed on said first end for shielding said urine sample application aperture;

c) a test strip comprising a dry porous carrier disposed within said hollow casing, wherein said dry porous carrier is nitrocellulose communicates indirectly with the exterior of said hollow casing via a bibulous urine sample receiving member which protrudes from said hollow casing and which acts as a reservoir from which said urine sample is released into said dry porous carrier,

said dry porous carrier having a detection zone;

said hollow casing including means permitting observation of said detection zone from outside said casing;

said dry porous carrier containing in a first zone upstream from said detection zone a labelled reagent comprising a colored particulate direct label conjugated to an anti-LH antibody, wherein said anti-LH antibody present in said labelled reagent is capable of specifically binding with said LH to form a first complex of said labelled reagent and said LH,

wherein said labelled reagent is dry on said test strip prior to use and is released into mobile form by said urine sample, said labelled reagent being freely mobile within said test strip when said test strip is wetted with said urine sample, said detection zone being spatially distinct from said first zone;

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

     1) coating at least said first zone with, or

     2) drying said labelled reagent onto a portion of said first 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 detection zone a means for specifically binding said first complex, said means for specifically binding said first complex being an unlabelled anti-LH antibody which is impregnated throughout the thickness of said dry porous carrier in said detection zone and is permanently immobilized in said detection zone.

wherein said anti-LH antibody of said labelled reagent and said unlabelled anti-LH antibody specifically bind to different LH epitopes,

said first zone and said detection zone being arranged such that said urine sample applied to said protruding bibulous urine sample receiving member and consequently to said dry porous carrier can migrate via said first zone into said detection zone,

said migration of said urine sample through said dry porous carrier conveying by capillarity said first complex to said detection zone whereat said means for specifically binding binds said first complex thereby to form a second complex;

said second complex being observable via said means permitting observation in said detection zone, thereby to indicate the presence of said LH in said urine sample.