Bi-directional lateral chromatographic test device

• US 5,006,474 A
• Filed: 12/16/1987
• Issued: 04/09/1991
• Est. Priority Date: 12/16/1987
• Status: Expired due to Term

• Current Assignee: Trinity Biotech
• Original Assignee: Disease Detection International, Inc. (MiCo IVD Holdings, LLC)
• Inventors: Whiteside, Julie, Horstman, Frederick C.
• Primary Examiner(s): Lacey, David L.
• Assistant Examiner(s): Waack, Janelle D.
• Application Number:
  US07/133,804
• Time in Patent Office: 1,210 Days
• Field of Search: 436/514, 436/524, 436/527, 436/170, 436/528, 436/539, 436/807, 436/808, 436/809, 436/825, 436/824, 436/161, 436/162, 436/515, 436/810, 422/61, 422/56, 422/57, 422/58, 422/60, 422/69, 422/70
• US Class Current: 436/524
• 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 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• 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 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 436/825 Pretreatment for removal of interfering factors from sample

Abstract

A chromatic test device for the performance of immuno- or chemical assays wherein a unitary planar fibrous filter body incorporates a sample application zone, a separation zone and a reaction zone and the application zone is in fluid communication with a first absorbent and the reaction zone is in fluid communication with a second absorbent to establish bilateral flow of the fluid component of the sample and of the analyte applied to the zones during the performance of an assay.

Claims

1. In a test device for performing solid phase immunoassays of a test sample having a liquid component, the combination of:

a planar fiber filter matrix which absorbs and conducts fluid, said matrix including a test sample application zone, a separation zone and a reaction zone, said separation zone separating said application zone from said reaction zone and being sufficiently long to inhibit migration of particulates in said sample into said reaction zone;

first absorbent material in communication with said sample application zone; and

second absorbent material in communication with said reaction zone so as to cause bilateral flow of the liquid component of said test sample;

said filter matrix at the location of communication between said sample application zone and said first absorbent material being constricted for causing said bilateral flow to be primarily from said application zone to said reaction zone, and secondarily from said application zone to said first absorbent material.

2. The device of claim 1 wherein said first absorbent material underlies said filter matrix.

3. The device of claim 1 in which said matrix is fabricated from a first material of a first porosity and said first and second absorbent materials are fabricated from a material of substantially different porosity from that of said first material.

4. The device of claim 1 in which said matrix has a range of pore sizes which prevent substantial migration of particulates in said sample from said absorption zone and said separation zone into said reaction zone, but permit bilateral flow of the liquid component of said test sample and any reagents accompanying the same into said reaction zone and permits bilateral flow of said fluid component and added reagents into said first and second absorbent materials.

5. The device of claim 4 in which said application zone is of bulbous configuration and said separation zone is constituted by an elongated shank portion integral with said application zone and with said reaction zone to permit fluid flow from said application zone through said separation zone into said reaction zone.

6. The device of claim 5, further including an extremity of said filter matrix overlying said first absorbent material so as to facilitate the aforesaid bilateral flow of said fluid component of said test sample.

27. The device of claim 1 in which such filter matrix includes a plurality of arms disposed in spaced relationship with one another, each of said arms including said test sample application zone, said separation zone and said reaction zone;

each of said arms being in communication with said first and second absorbent

7. In a test device for performing solid phase immunoassays, the combination of:

a housing including a cover;

first and second apertures through said cover;

a planar filter matrix located in said housing under said cover, said filter matrix incorporating a test sample application zone at said first aperture, a separation zone in communication with said test sample application zone and a reaction zone at said second aperture in communication with said separation zone;

first absorbent material located in said housing in fluid communication with said application zone; and

second absorbent material in said housing in spaced relationship with said first absorbent material and in fluid communication with said reaction zone so as to cause bilateral flow of the fluid component of said test sample;

said first aperture and said application zone being disposed between said first absorbent material and said reaction zone; and

said second aperture and said reaction zone being disposed between said application zone and said second absorbent material.

8. The test device of claim 7 in which the fluid communication of said application zone with said first absorbent means is defined by said first absorbent means being disposed contiguous to a portion of the perimeter of said application zone.

9. The device of claim 7 in which said fluid communication between said first absorbent material and said application zone is defined by said first absorbent material being disposed below a portion of said filter matrix.

10. A method of performing an immunoassay in conjunction with a chromatographic test device which incorporates a planar filter matrix having sample application separation and reaction zones and first and second absorbent means in fluid communication respectively with said application and reaction zones so as to cause bilateral flow of fluids applied to said zones, comprising the steps of:

applying a suitaBle volume of liquid sample containing particulates to said sample application zone;

applying a suitable volume of wash reagent to said sample application zone;

where after each of said sample and wash applications capillary forces within said filter matrix bilaterally draw the fluid portion of said sample primarily in the direction of said separation zone, but secondarily in the direction of said first absorbent means;

and where said application and separation zones entrap said particulates;

and where the fluid portion of said sample is drawn into said reaction zone which is treated with a reagent which reacts and binds with ananlyte component being assayed in said sample, said reagent being immobilized to the fiber matrix.

applying a suitable volume of wash reagent to said reaction zone to wash away unreacted sample components under said bilateral flow away from the reaction zone and toward the separation and sample application zones;

applying a tracer reagent complementary to said analyte component being assayed to said reaction zone, said tracer reagent being a reagent conjugated with a tracer;

washing said reaction zone; and

applying chromatic eliciting substrate to said reaction zone to produce a chromatic reaction determining the presence of said tracer in said reaction zone.

11. The method of claim 10 in which said tracer is an enzyme.

12. The method of claim 10 in which said tracer is a radionuclide.

13. The method of claim 10 in which said tracer is a fluorescent dye.

14. In a test device for performing solid phase immunoassays, the combination of:

a planar fiber filter matrix, said matrix including a test sample application zone, a separation zone and a reaction zone, said separation zone separating said application zone from said reaction zone and being sufficiently long to inhibit migration of particulates in said sample into said reaction zone;

first absorbent material in communication with said sample application zone, and second absorbent material in communication with said reaction zone so as to cause bilateral flow of the liquid component of said test sample;

said application zone being of bulbous configuration and said separation zone being an elongated relatively narrow shank portion integral with said application zone and with said reaction zone to permit fluid flow from said application zone through said separation zone into said reaction zone.

15. In a test device for performing solid phase immunoassays, the combination of:

a planar fiber filter matrix, said matrix including a test sample application zone, a separation zone and a reaction zone, said separation zone separating said application zone from said reaction zone and being sufficiently long to inhibit migration of particulates in said sample into said reaction zone;

first absorption means in communication with said sample application zone, and second absorption means in communication with said reaction zone so as to cause bilateral flow of the liquid component of said test sample;

the structure of said filter matrix at the location of said communic causing the bilateral flow to be primarily in the direction from said application zone to said reaction zone and secondarily in the opposite direction;

said first and second absorbent means being of substantially larger cross section than said separation zone.

16. A method of performing an assay for an analyte in a sample fluid comprising:

immobilizing a specific reagent complimentary to said analyte to a reaction zone of a fiber filter matrix, said fiber filter matrix also having a sample application and separation zones, and first and second absorbent means in fluid communication, respectively, with said application and reaction zones so as to cause bilateral flow of fluids applied to said sample application zone.

applying a volume of said sample fluid to said sample application zone.

where said fiber matrix draws the fluid portion of said sample primarily in the direction fo the separation zone, but secondarily in the opposite direction under said bilateral flow;

and where said fibermatrix draws some of said fluid portion of said sample into said reaction zone;

and also where any analytes contained within said fluid drawn into said reaction zone react and bind with said specific reagent;

washing said reaction zone of unreacted sample components;

applying a tracer reagent which binds to said analyte component being assayed to said reaction zone said tracer reagent being a reagent conjugated with a tracer;

washing said reaction zone of unbound tracer reagent; and

determining the presence of said analyte in said sample by the presence or absence of said tracer in said reaction zone.

17. The method of claim 16, additionally comprising applying a blocking agent in order to decrease non-specific binding of extraneous components of said sample after the step of immobilizing said reagent onto said reaction zone.

18. The method of claim 16, wherein said sample is a bodily fluid from a mammal.

19. The method of claim 18, wherein said bodily fluid is selected from the group consisting of blood and urine.

20. The method of claim 16, wherein said immobilized reagent is an antibody specific to said analyte, wherein said tracer reagent is a tracer conjugated to an antibody specific to said analyte, and wherein the presence of said tracer upon the completion of the assay indicates the presence of said analyte in said sample.

21. The method of claim 16, wherein said immobilized reagent is an antibody specific to said analyte, wherein said tracer reagent is a tracer conjugated to a molecule having an antigenic site reactive with said antibody, and wherein the absence of said tracer upon the completion of the assay indicates the presence of said analyte in said sample.

22. The method of claim 16, wherein said immobilized reagent is an antigen specific to a first antibody, said first antibody to be detected in said sample fluid, wherein said tracer reagent is a tracer conjugated to a second antibody, said second antibody being specific to said first antibody, and wherein the presence of said tracer upon the completion of the assay indicates the presence of said first antibody in said sample.

23. The method of claim 16, wherein said immobilized reagent is an antigen specific to a first antibody, said first antibody to be detected in said sample fluid, wherein said tracer reagent is a tracer conjugated to a second antibody, said second antibody being specific to said antigen, and wherein the absence of said tracer upon the completion of the assay indicates the presence of said first antibody in said sample.

24. A method of performing an immunoassay in conjuncation with a chromatographic test device which incorporates a planar filter matrix having sample application separation and reactionzones, and first and second absorbent means in fluid communication, respectively, with said with said application and reaction zones so as to cause bilateral flow of fluids applied to said zones, comprising the steps of:

applying a suitable volume of said sample fluid to said sample application zone;

where capillary forces within said fiber matrix draw the fluid portion of said sample primarily in the direction of said separation zone, but secondarily in the direction of said first absorbent means under said bilateral flow;

and where said application and separation zones entrap said particulates;

and also where the fluid portion of said sample is drawn into said reaction zone which is treated with a reagent which reacts and hinds with analyte component being assayed in said sample said reagent being immobilized to the fiber matrix;

washing said reaction zone to wash away unreacted sample components away from the reaction zone;

applying a tracer reagent complementary to said analyze component being assayed to said reaction zone, said tracer reagent being a reagent conjugated with a tracer;

washing said reaction zone, and determining the presence of said tracer in said reaction zone.

25. The method of claim 24, wherein the step of determining the presence of said tracer comprises applying chromic-eliciting substrate to said reaction zone, said substrate being acted upon by said tracer to produce a chromatic reaction.

26. The method of claim 24, additionally comprising applying a suitable volume of wash reagent to said sample application zone after the step of applying a suitable volume of liquid sample to said sample application zone.