Competitive ELISA for the detection of HTLV-III antibodies

• US 4,661,445 A
• Filed: 05/24/1985
• Issued: 04/28/1987
• Est. Priority Date: 05/24/1985
• Status: Expired due to Fees

• Current Assignee: United States Department Of Health And Human Services (Government of the United States of America)
• Original Assignee: United States Department Of Health And Human Services (Government of the United States of America)
• Inventors: Gallo, Robert C., Saxinger, W. Carl
• Primary Examiner(s): Nucker, Christine M.
• Assistant Examiner(s): Benson, Robert
• Application Number:
  US06/737,458
• Time in Patent Office: 704 Days
• Field of Search: 435/5, 435/7, 435/810, 436/518, 436/528, 436/543, 436/808, 436/809, 436/811, 436/825, 935/110
• US Class Current: 435/5
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: G01N 33/56988 HIV or HTLV
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 435/974 Aids related test
• CPC Class Codes: Y10S 435/975 Kit
• CPC Class Codes: Y10S 436/808 Automated or kit
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays
• CPC Class Codes: Y10S 436/811 Test for named disease, body condition or organ function
• CPC Class Codes: Y10S 436/825 Pretreatment for removal of interfering factors from sample

Abstract

A competitive enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies is disclosed. This ELISA technique is more sensitive, more specific, and more accurate than known ELISA techniques. The competitive ELISA of this invention is particularly suited to the detection of human T-cell leukemia-lymphoma virus type III (HTLV-III). Antigenic or viral fragments are bound to a solid support. The bound fragments are then incubated with a heterologous antiserum which binds to the bound fragments. Test sera is then added to the bound fragments and tested for absorbance.

Claims

1. A method for the detection of HTLV-III antibodies in a test sample comprising

coating microtiter plates with purified HTLV-III antigen to form antigen-coated wells;

incubating a portion of said antigen coated wells with heterologous anti-HTLV-III;

adding test sera to said wells;

screening said wells for the amount of binding of HTLV-III antibodies by comparing said wells containing said heterologous anti-HTLV-III to control wells, said control wells being incubated without said heterologous anti-THLV-III.

2. A test kit for the detection of HTLV-III antibodies comprising in combination

an insoluble surface or support containing microtiter wells;

HTLV-III fragments which are bound to said support; and

heterologous antiserum which specifically binds to said HTLV-III fragments; and

means for detecting the amount of antibodies in a test sample which bind to said HTLV-III fragments.