The reactivity of sera from patients of Tsutsugamushi disease (scrub typhus) with the antigenic polypeptides of Rickettsia tsutsugamushi was analyzed by the immunoblotting method. The reactivity varied greatly among the sera of individual patients tested. IgG and IgM antibodies of most patients reacted with the 54-56 kilodalton (54-56K) polypeptide located on the rickettsial surface, suggesting that this polypeptide is a predominant antigen in the infection. Other polypeptides of 60K, 50-52K, 46-47K, 35K, and 21-25K were reactive with some but not all sera. From the reactivity of these polypeptides, it was suggested that the 54-56K polypeptide is both strain-specific and group-specific, the 60K polypeptide is group-specific, and the 35K and 21-25K polypeptides are subgroup-specific. IgG antibodies seem to be more cross-reactive with polypeptides of multiple strains than IgM antibodies and have a tendency of increased cross-reactivity that was observed in the sera obtained at the later stage of illness.