A 4-layer modification of ELISA for the determination of toxoplasma antibodies is described. In 103 Finnish blood donors 27 had antibodies against Toxoplasma gondii. One donor had IgM antibodies and IgA antibodies were found in 9. In patients with acute toxoplasmosis a vigorous IgG antibody response resulted in high antibody levels soon after infection, declining gradually to mean adult levels in approximately 2 yr. IgM antibodies appeared during the earliest phases of infection and disappeared as early as in 1 or 2 months in some cases and in most cases by the 6th month after infection. An IgA antibody response was also always seen. It was slower than the IgM response and could therefore be used to improve the laboratory diagnosis especially in cases where IgM antibodies had already disappeared and no further increase in IgG antibodies could be detected.