Immune Complex Transfer Enzyme Immunoassay for Antibodies Using Recombinant Proteins Labeled with 2,4-Dinitrophenyl Group and β-D-Galactosidase
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Ultrasensitive enzyme immunoassays (immune complex transfer enzyme immunoassays) were developed for antibody IgGs. Antibody IgGs were reacted with 2, 4-dinitrophenyl-antigen and antigen-β-<SUB>D</SUB>-galactosidase conjugate, and the immune complexes comprising the three components were trapped onto solid phase coated with (anti-2, 4-dinitrophenyl group) IgG and were transferred to solid phase coated with (antihuman IgG γ-chain) IgG in the presence of ε<I>N</I>-2, 4-dinitrophenyl-<SUB>L</SUB>-lysine. These immunoassays were much more sensitive and specific than currently available conventional methods and were applied for detection of antibody IgGs to HIV-1. Diagnosis and confirmation of HIV-1 infection using urine, whole saliva and serum samples became more reliable with less indeterminate results. And the window period in diagnosis of HIV-1 infection, during which diagnosis of HIV-1 infection is not possible due to the absence of detectable antibodies to HIV-1, was shortened by approximately two weeks with the immune complex transfer enzyme immunoassay for simultaneous detection of p24 antigen of HIV-1 and antibody IgGs to p17 antigen and reverse transcriptase of HIV-1 in a single assay tube. Furthermore, it has recently become possible to perform the immune complex transfer enzyme immunoassay within 2 to 3 hr with higher sensitivity and, as a result, to further shorten the window period in diagnosis of HIV-1 infection.
- ACTA HISTOCHEMICA ET CYTOCHEMICA
ACTA HISTOCHEMICA ET CYTOCHEMICA 29(4), 273-281, 1996-08-01
JAPAN SOCIETY OF HISTOCHEMISTRY AND CYTOCHEMISTRY