Whole blood assays using porous membrane support devices

• US 5,096,809 A
• Filed: 07/25/1988
• Issued: 03/17/1992
• Est. Priority Date: 07/25/1988
• Status: Expired

• Current Assignee: Quidel Corporation
• Original Assignee: Pacific Biotech, Inc. (Quidel Corporation)
• Inventors: Fan, Eugene, Chen, Fon-Chiu M.
• Primary Examiner(s): Nucker, Christine
• Assistant Examiner(s): Stucker, Jeffrey
• Application Number:
  US07/223,520
• Time in Patent Office: 1,331 Days
• Field of Search: 435/6, 435/7, 435/28, 422/55, 422/56, 436/67, 436/178, 436/514, 436/535, 210/789
• US Class Current: 435/7.9
• 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 33/54391 based on vertical flow
• CPC Class Codes: Y10T 436/255 including use of a solid sorbent, semipermeable membrane, or liquid extraction

Abstract

Disclosed is a method for detecting analytes in samples of whole blood using solid-phase, permeable support assay devices. The whole blood assays require no steps or reagents other than those necessary to carry out the same analysis on plasma or serum using the devices. The color visualization of the analytical field on which the interpretation of the assay depends is not affected by the presence of intact erythrocytes or any degree of hemolysis in the blood sample.

Claims

1. A method for analyzing whole blood for an analyte recognized by an analyte specific reagent, comprising the steps of:

applying whole blood to a liquid permeable support layer of an assay device, wherein said layer has said analyte specific reagent fixed thereto prior to said introduction of said blood, and wherein said whole blood is applied to said layer;

drawing colored components of said blood through said layer; and

forming a color on said layer in response to interaction between said analyte and said analyte specific reagent when analyte is present in said blood, thereby visually indicating on said layer the presence or absence of said analyte in said blood.

2. The method of claim 1, further comprising the step of:

lysing red blood cells to release colored components thereof and to permit said components to be drawn through said layer.

3. The method of claim 2, wherein said lysing step is performed prior to said applying step.

4. The method of claim 2, wherein said lysing step occurs after said applying step.

5. The method of any one of claims 1-4, wherein said layer is a membrane and wherein said drawing step comprises wicking liquid components of said blood away from said membrane.

6. The method of claim 5, further comprising the steps of:

adding a liquid to said layer after said applying step, which liquid is relatively colorless in comparison to said colored components of said blood; and

drawing said relatively colorless liquid, together with said colored components, through said layer to enhance the visibility of color formed on said layer in said forming step.

7. The method of claim 5, wherein the membrane has an average pore size no greater than about 25 μ

m.

8. The method of claim 5, wherein the membrane has an average pore size no greater than about 10 μ

m.

9. The method of claim 5, wherein the membrane has an average pore size no greater than about 5 μ

m.

10. The method of claim 5, wherein said analyte comprises one member of a ligand-antiligand pair and said analyte specific reagent comprises the other member of said ligand-antiligand pair.

11. The method of claim 10, wherein said ligand-antiligand pair comprises an antigen and an antibody.

12. The method of claim 1, wherein said color change occurs as a result of the action of an enzyme on a substrate, which enzyme is immobilized on said membrane at the time of said color change.

13. The method of claim 1, wherein said analyte and said analyte specific reagent together comprise an enzyme-substrate pair.

14. The method of claim 1, wherein said drawing step comprises drawing hemoglobin from lysed red blood cells through said layer.

15. The method of claim 1, wherein said drawing step comprises drawing whole red blood cells through said layer.

16. The method of claim 1-4, comprising an assay for infectious mononucleosis.

17. The method of claim 16, wherein the analyte specific reagent is mononucleosis antigen and said analyte is infectious mononucleosis heterophile antibody.