A one day enzyme immunoassay for the detection of rheumatoid factors of the IgG, IgM, and IgA class is described. The assay utilizes rabbit IgG as solid-phase reactant and the biotin-avidin interaction for the coupling of enzyme to indicator antibody. Three different indicator antibodies discriminated effectively between rheumatoid arthritis patients and normal subjects. F(ab')2 fragments of goat antibodies were found best suited for the test. Rheumatoid factor activity was expressed in U/ml by comparing samples to an internal standard, which was related to the WHO international reference serum for rheumatoid arthritis. Rheumatoid factor activity (U/ml) in the IgM-specific assay showed a close correlation to the latex agglutination titer. Avidity indices estimated from the slopes of the dose response curves of test sera were significantly higher for rheumatoid arthritis patients than for a group of healthy persons, indicating a higher avidity of the rheumatoid factors in the patients' sera.