We developed an enzyme immunoassay (EIA) system to detect antibodies to human T-lymphotropic virus type I (HTLV-I). This system uses chemically synthesized oligopeptides to capture anti-HTLV-I antibodies in serum. The two epitopes of HTLV-I proteins exhibiting the most specific antigen-antibody reaction reside within amino acids 100-130 of p19, a core protein encoded by gag, and amino acids 175-199 of gp46, an envelope glycoprotein encoded by env. This new assay uses synthetic peptides corresponding to these two regions modified by adding two lysine residues at the amino terminal of each peptide to facilitate the binding to the surface of the microtiter plate wells. We compared the performance of our EIA with gelatin-particle-agglutination (PA) and indirect-immunofluorescence (IF) assays, both of which use viral proteins purified from virus-carrying cell cultures. Mass screening by EIA with various synthetic peptides was more accurate than the current confirmatory IF assay.