Anti-HTLV-I IgG in urine detected by sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using a synthetic peptide, Cys-env gp46(188-224), as antigen

✍ Scribed by Seiichi Hashida; Kouichi Hirota; Takeyuki Kohno; Dr. Eiji Ishikawa

• Publisher: John Wiley and Sons
• Year: 1994
• Tongue: English
• Weight: 610 KB
• Volume: 8
• Category: Article
• ISSN: 0887-8013
• DOI: 10.1002/jcla.1860080307

✦ Synopsis

Abstract

Antibody IgG to human T‐cell leukemia virus type I (HTLV‐I) in urine was detected by a sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using a synthetic peptide, Cys‐env gp46(188–224), as antigen, the sensitivity and specificity of which were 100 and 98.5%, respectively, using serum samples. Anti‐HTLV‐I IgG in urine was reacted simultaneously with 2,4‐dinitrophenyl‐bovine serum albumin–Cys‐env gp46(188–224) conjugate and Cys‐env gp46(188–224)‐β‐D‐galactosidase (Escherichia coli) conjugate. The complex formed, consisting of the three components, was trapped onto polystyrene balls coated with affinity‐purified (anti‐2,4‐dinitrophenyl group) IgG, eluted with ϵ__N__‐2,4‐dinitrophenyl‐L‐lysine and transferred to polystyrene balls coated with affinity‐purified (anti‐human IgG γ‐chain) IgG. Finally, bound β‐D‐galactosidase activity was assayed by fluorometry. Thirty‐one urine samples from seropositive subjects and 100 urine samples from seronegative subjects were tested. The sensitivity and specificity were 87 and 100%, respectively, with unconcentrated urine samples and 94 and 100%, respectively, with approximately 10‐fold concentrated urine samples. These results were superior to those by the conventional ELISA and gelatin particle agglutination test. © 1994 Wiley‐Liss, Inc.