The sensitivity and specificity of an indirect immunofluorescence assay (IFA) and ELISA for Lyme disease were estimated. Sera from patients with Lyme disease, patients with other infections, and healthy individuals were examined. Significant cross-reactivity occurred only with sera from patients with syphilis, yaws, and pinta . All tested sera from patients with Lyme disease, however, gave negative results in the rapid reagin screening test and the microhemagglutination assay for antibodies to Treponema pallidum confirmatory for syphilis. When sera from patients with treponemal diseases were excluded from the analysis, the IFA and ELISA were highly specific, having 97% and 100% reliability, respectively. Sensitivity of both tests varied with the stage of disease but was 100% for both tests during complicated Lyme disease. The results indicate that both tests are highly specific and sensitive for complicated Lyme disease but relatively insensitive for patients with erythema chronicum migrans alone.