The seroprevalence of HTLV-1 antibodies was investigated in 405 serum samples from healthy South American Indians (Mapuches) from Chile, using enzyme linked immunoassay (ELISA), Western immunoblot (WB) and radioimmuno precipitation assay (RIPA). Six samples were positive by ELISA; 3 of them were confirmed by WB/RIPA. Thus, we observed a seroprevalence of 0.7% for HTLV-1 antibodies in healthy Mapuches.