Enzyme-linked immunosorbent assay that uses labeled antigen for detection of immunoglobulin M and A antibodies in toxoplasmosis: comparison with indirect immunofluorescence and double-sandwich enzyme-linked immunosorbent assay

Abstract

A direct enzyme-linked immunosorbent assay (ELISA) is described that uses horseradish peroxidase-labeled antigen for detection of immunoglobulin M (IgM) and IgA antibodies to toxoplasma. In this assay, polystyrene microtiter plates were sensitized with anti-human IgM or IgA antibody to separate IgM or IgA from other classes of antibody. The presence of IgM or IgA antibodies to toxoplasma (Tox-IgM, Tox-IgA) was then detected by sequential addition of soluble horseradish peroxidase-labeled toxoplasma antigen and substrate. As judged by examining sucrose gradient-fractionated sera, the assay was specific for IgM or IgA classes of antibody. In contrast to the indirect immunofluorescence for IgM antibodies to toxoplasma, no inhibition of IgM reactivity by specific IgG antibodies could be detected. Furthermore, rheumatoid factor did not cause false-positive results. Of 80 single sera with high antibody titer to toxoplasma in indirect immunofluorescence and complement fixation, 40 were positive in the direct ELISA for Tox-IgM, 36 were positive in the double-sandwich ELISA, and only 21 were positive in the indirect immunofluorescence for Tox-IgM when whole serum was used. In the indirect immunofluorescence, another 13 sera became positive after sucrose gradient fractionation. The direct ELISA for IgA antibodies to toxoplasma was positive in 43 sera, of which 39 were positive in the direct ELISA for Tox-IgM. High levels of IgM antibodies were found within 3 months after the onset of symptoms, slowly decreasing thereafter. Tox-IgM may persist for more than 1 year after infection.