Detection of multiple analytes from a single sample using a multi-well, multi-analyte flow-through diagnostic test device

• US 7,052,831 B2
• Filed: 12/08/2000
• Issued: 05/30/2006
• Est. Priority Date: 09/29/2000
• Status: Expired due to Fees

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Fletcher, Robert C., Ozinskas, Alvydas J., Kopher, Kenneth, Gill, John
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US09/732,838
• Publication Number:
  US 20020115062A1
• Time in Patent Office: 1,999 Days
• Field of Search: 422/56, 422/57, 422/58, 422/61, 435/287.1, 435/287.2, 435/5, 435/7.9, 435/7.92, 435/975, 435/973, 436/518, 436/809
• US Class Current: 435/5
• CPC Class Codes: G01N 33/54386 Analytical elements
• CPC Class Codes: G01N 33/56983 Viruses
• CPC Class Codes: Y10S 435/973 Simultaneous determination of more than one analyte
• CPC Class Codes: Y10S 435/975 Kit
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays

Abstract

The present invention provides a method and device for conducting a rapid in vitro enzyme immunoassay test for the direct and qualitative detection of two or more viral antigens from specimens of symptomatic patients. The method for immunoassay of viral antigens is performed on a membrane. Non-immunological capture of viral antigens takes place by absorption onto the membrane. Captured antigen binds to a detection reagent that includes a label conjugated to a specific antibody. The test is a differentiated test such that two or more viral antigens may be distinguished from each other in a single test. The invention also includes a kit for performing an assay in accordance with the method of the present invention, wherein the kit comprises the device of the present invention.

Claims

1. A kit for performing a flow-through analytical test for detecting the presence of and distinguishing between Influenza A viral antigens and Influenza B viral antigens in a biological sample suspected of containing one or both of Influenza A viral antigens and Influenza B viral antigens, the kit comprising:

(a) an analytical test device having a plurality of wells, wherein the wells do not communicate with each other; and

wherein each well comprises;

a filter stack, the filter stack comprising;

(i) a porous membrane having an upper surface and a lower surface; and

(ii) an absorbent material, wherein said lower surface of said porous membrane and said absorbent material are in physical contact and in fluid communication, and wherein upon contact of a biological fluid sample suspected of containing one or both of the Influenza A viral antigens and the Influenza B viral antigens with said porous membrane, said fluid is able to flow through said membrane into said absorbent material, such that at least a portion of said viral antigens present in said biological fluid sample bind to said porous membrane;

(b) a first detection reagent capable of recognizing Influenza A viral antigens;

(c) a second detection reagent capable of recognizing Influenza B viral antigens; and

(d) a reagent capable of generating a signal upon detection of one or both of Influenza A viral antigens and Influenza B viral antigens.

2. The kit of claim 1, wherein the test device comprises two wells.

3. The kit of claim 1, wherein the first detection reagent comprises an anti-Influenza A monoclonal murine [antibodies (2)] antibody-enzyme conjugate.

4. The kit of claim 1, wherein the second detection reagent comprises an anti-Influenza B monoclonal murine antibody-enzyme conjugate.

5. The kit of claim 1, wherein the reagent capable of generating a signal comprises chromogen.

6. The kit of claim 1, further comprising an extraction reagent.

7. The kit of claim 6, wherein the extraction reagent comprises a mucolytic agent.

8. The kit of claim 1, further comprising at least one wash reagent.

9. The kit of claim 1, further comprising a stop reagent.

10. The kit of claim 9, wherein the stop reagent comprises citric acid.