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A direct antibody capture enzyme immunoassay (EIA) for the detection of rubella-specific immunoglobulin M (IgM) antibody was developed. Polystyrene microtiter strips coated with rabbit anti-human IgM were used as solid phase, and a semipurified rubella antigen labeled with horseradish peroxidase was used as indicator. By testing a panel of 238 serum specimens (not including sera from newborns with congenital rubella), the direct EIA was compared with the indirect EIA used routinely in our diagnostic unit and with a commercial IgM capture EIA (RubEnz M II) that employs a horseradish peroxidase-labeled anti-rubella monoclonal antibody as indicator. Overall agreement of direct EIA with indirect EIA and RubEnz M II was 95%, whereas agreement between direct and indirect EIA was 96.2%, and agreement between direct EIA and RubEnz M II was 97.8%. Sensitivity of the direct assay was higher than that of the indirect EIA and similar to that of the RubEnz M II assay. Specific IgM antibody could always be detected in serum specimens taken from patients with primary acute rubella infection between days 4 and 56 after the onset of rash. The assay was highly specific, and it was not affected either by rheumatoid factor or by high levels of specific IgG in sera. Another important advantage that the direct EIA has over the indirect EIA is that it requires 10 to 20 times less antigen per serum specimen tested.

Detection of specific immunoglobulin M antibody to rubella virus by use of an enzyme-labeled antigen