Sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for (antihuman T-cell leukemia virus type I) immunoglobulin G in serum using a synthetic peptide, Ala-Cys-Env gp46(237–262), as antigen

✍ Scribed by Takeyuki Kohno; Iwane Sakoda; Dr. Eiji Ishikawa

• Publisher: John Wiley and Sons
• Year: 1992
• Tongue: English
• Weight: 714 KB
• Volume: 6
• Category: Article
• ISSN: 0887-8013
• DOI: 10.1002/jcla.1860060311

✦ Synopsis

A sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for (antihuman T-cell leukemia virus type I) IgG (anti-HTLV-l IgG) in serum using a synthetic peptide, Ala-Cys-env gp46(237-262), of HTLV-I is described. Anti-HTLV-l IgG in test serum, which had been incubated with excess of inactive p-D-galactosidase to eliminate interference by anti-p-D-galactosidase antibodies, was reacted simultaneously with 2,4-dinitrophenyl-bovine serum albumin-Ala-Cys-env gp46(237-262) conjugate and Ala-Cys-env gp46(237-262)-p-D-galactosidase conjugate. The complex formed consisting of the three components was trapped onto polystyrene balls coated with affinity-purified (anti-2,4dinitrophenyl group) IgG. After washing to eliminate nonspecific IgG in the test serum and excess of the p-D-galactosidase conjugate, the complex was eluted from the polystyrene balls with ~N-2,4-dinitrophenyl-L-lysine and transferred to polystyrene balls coated with affinity-purified (anti-human IgG y-chain) IgG. p-0-galactosidase activity bound to the (antihuman IgG y-chain) IgG-coated polystyrene balls was assayed by fluorometry. This assay was more sensitive than other methods using HTLV-I as antigen, and most negative and positive sera were discriminated. However, some results appeared to be false positive or false negative, and the peptide, Ala-Cysenv gp46(237-262), was suggested to be useful, in combination with other peptides, for improving the reliability of serodiagnosis by separately demonstrating antibodies against as many different epitopes of HTLV-I as possible. 01992 ~itey-Liss, Inc.