Capillary immunoassay and device therefor comprising mobilizable particulate labelled reagents

• US 5,622,871 A
• Filed: 07/15/1993
• Issued: 04/22/1997
• 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: May, Keith, Prior, Michael E., Richards, Ian
• Primary Examiner(s): Spiegel, Carol A.
• Application Number:
  US08/102,313
• Time in Patent Office: 1,377 Days
• Field of Search: 722/56-58, 436/501, 436/530, 436/810, 436/814, 436/514, 436/515, 436/518, 436/523, 436/524, 436/541, 436/510, 436/538, 436/817, 436/818, 435/810, 435/7.92-7.95, 435/970, 422/60, 427/2, 427/2.11
• 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, 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, 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 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 liquid biological sample application aperture and means permitting observation of a test result;

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 liquid biological sample, said carrier having a test result zone observable via said means permitting observation, said test strip, in the dry unused state, containing 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 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 result zone in the presence of, a material comprising a 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 via said means permitting observation, thereby to indicate the presence of said analyte in said liquid biological sample.

2. The device according to claim 1, wherein said label is selected from the group consisting of dye sols and metallic sols.

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

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

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

6. The test device according to claim 5, wherein said nitrocellulose has a pore size greater than about 5 microns.

7. The test device according to claim 5, wherein said nitrocellulose has a pore size of about 8-12 microns.

8. The test device according to claim 1 which further comprises a control zone downstream from said test result zone in said dry porous carrier to indicate that said liquid biological sample is conveyed by capillarity beyond said test result zone, and a control zone observation aperture in said casing, said control zone also being observable from outside said hollow casing through said control zone observation aperture.

9. The test device according to claim 8, wherein said control zone contains a means for binding said labelled reagent which comprises specific binding means and is immobilized in said control zone.

10. 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 labelled reagent not bound to said analyte washed out of said test zone.

11. The test device according to claim 1, wherein said means for binding is impregnated throughout said dry porous carrier in said test zone.

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

13. The test device as in claim 1, wherein said means permitting observation comprises a test result observation aperture defined through a wall of said casing.

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

15. The test device according to claim 14, wherein said protein is bovine serum albumin.

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

17. 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.

18. 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.

19. The test device according to claim 18, wherein said protein is selected from the group consisting of bovine serum albumin and milk protein.

20. An analytical method comprising:

contacting an analytical test device for detecting an analyte suspected of being present in a liquid biological sample, said device including;

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

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 liquid biological sample, said carrier having a test result zone observable via said means permitting observation, said test strip, in the dry unused state, containing 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 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 result zone in the presence of, a material comprising a 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 via said means permitting observation, thereby to indicate the presence of said analyte in said liquid biological sample, with said liquid biological sample suspected of containing said analyte;

allowing said liquid biological sample to convey by capillary action through said dry porous carrier into said test result zone and said labelled reagent is conveyed therewith;

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

21. The method according to claim 20, wherein said dry porous carrier is a strip of dry porous material, and has a control zone downstream from said test result zone, to indicate to a user that the test has been completed;

and said hollow casing has a second observation aperture through which said control zone is visible and wherein said liquid biological sample is a urine sample whereby urine is conveyed by capillarity through said dry porous carrier into said test result zone and said control zone, and said test result is obtained by observing the extent to which said labelled reagent-analyte complex becomes bound in said test result zone.

22. The method according to claim 21, wherein said control zone contains a means for binding said labelled reagent which comprises specific binding means and is immobilized in said control zone.

23. The method according to claim 20, wherein said material further comprises a protein.

24. The method according to claim 23, wherein said protein is bovine serum albumin.

25. The method according to claim 20, wherein said sugar is selected from the group consisting of sucrose, lactose and dextran.

26. The method according to claim 20, 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.

27. The method according to claim 20, 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.

28. The method according to claim 27, wherein said protein is selected from the group consisting of bovine serum albumin and milk protein.

29. 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 aperture, a test result observation aperture, and a control zone 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 aperture to receive applied liquid biological sample, said carrier having a test result zone observable through said test result observation aperture and a control zone observable through said control zone observation aperture, said test strip, in the dry unused state, containing 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 applied liquid biological sample;

c) 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 to said analyte to wash out of said test zone;

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

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 result zone in the presence of, a material comprising a sugar, in an amount effective to reduce interaction between said test strip and said labelled reagent;

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 means for binding said first complex 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.

30. The test device according to claim 29, wherein said control zone contains a means for binding said labelled reagent which comprises specific binding means and is immobilized in said control zone.

31. The method according to claim 29, wherein said material further comprises a protein.

32. The method according to claim 31, wherein said protein is bovine serum albumin.

33. The test device according to claim 29, wherein said sugar is selected from the group consisting of sucrose, lactose and dextran.

34. The test device according to claim 29, 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.

35. The test device according to claim 29, 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.

36. The test device according to claim 35, wherein said protein is selected from the group consisting of bovine serum albumin and milk protein.