Open AccessImmune complex transfer enzyme immunoassay for antibody IgM to HIV‐1 p17 antigen
Author(s) -
Hashida Seiichi,
Ishikawa Setsuko,
Nishikata Ichiro,
Hashinaka Kazuya,
Oka Shinichi,
Ishikawa Eiji
Publication year - 1998
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/(sici)1098-2825(1998)12:6<329::aid-jcla1>3.0.co;2-h
Subject(s) - immune complex , antigen , antibody , immunoassay , seroconversion , immunoglobulin m , immune system , immunoglobulin g , bovine serum albumin , microbiology and biotechnology , hiv antigens , virology , chemistry , biology , immunology , virus , viral disease
The immune complex transfer enzyme immunoassay for antibody IgM to HIV‐1 p17 antigen is described. Serum samples containing antibody IgM to HIV‐1 p17 antigen were incubated simultaneously with 2,4‐dinitrophenyl‐bovine serum albumin‐recombinant p17 (rp17) conjugate and rp17‐β‐ D ‐galactosidase conjugate, and the immune complex formed comprising the three components was trapped onto colored polystyrene beads coated with affinity‐purified (anti‐2,4‐dinitrophenyl group) IgG. Subsequently, the immune complex was transferred to white polystyrene beads coated with monoclonal mouse (antihuman IgM) IgG in the presence of excess of ϵN‐2,4‐dinitrophenyl‐ L ‐lysine. The signal for antibody IgM to p17 antigen was the fluorescence intensity by fluorometric assay of β‐ D ‐galactosidase activity bound to the white polystyrene beads. The periods of time required for the formation, trapping, and transferring of the immune complex comprising the three components were more than 4 hr, 2 hr, and 3 hr, respectively. The immunoassay developed was shown to be specific by inhibition of transferring the immune complex in the presence of excess of nonspecific IgM but not IgG. Signals for antibody IgM to p17 antigen with serum samples of HIV‐1 seroconversion serum panels,—that is, with serum samples in early stages of the infection—tended to be higher than those with serum samples from HIV‐1 asymptomatic carriers probably long after the infection and patients with ARC and AIDS. In contrast, signals for antibody IgG to p17 antigen with serum samples of HIV‐1 seroconversion serum panels tended to be higher than signals for antibody IgM to p17 antigen but were much lower than signals for antibody IgG to p17 antigen with serum samples from HIV‐1 asymptomatic carriers and patients with ARC and AIDS. J. Clin. Lab. Anal. 12:329–336, 1998. © 1998 Wiley‐Liss, Inc.