An 80-kilodalton Trypanosoma cruzi antigen is eliminated in the urine of infected hosts during the acute stage of Chagas' disease. We show that affinity-purified urinary antigen is recognised by IgM antibodies in the sera from acute chagasic patients. Comparing our urinary antigen assay with that using a whole T. cruzi lysate antigen for IgM antibody detection, we demonstrated that ELISA with urinary antigen increases the diagnostic sensitivity and specificity of IgM serology in recent chagasic infection. Twenty-six of 30 patients with acute T. cruzi infection had serum IgM antibodies that reacted with urinary antigen by ELISA, while lysate antigen IgM was detected in 24 sera. When sera from patients suffering other parasitoses were tested, strong cross-reactions occurred in ELISA with T. cruzi lysate antigen, whereas ELISA with urinary antigen proved to better discriminate acute chagasic patients. Human antibodies to urinary antigen immunoprecipitated this T. cruzi urinary antigen and also inhibited the binding of monoclonal antibody to urinary antigen in an inhibition assay. These findings suggest that urinary antigen may be useful for the development of serodiagnostic procedures for acute T. cruzi infection.