Antigenicity of adult Opisthorchis viverrini, metacercariae and eggs was investigated by radio-immunoprecipitation and polyacrylamide gel electrophoresis using sera from patients with opisthorchiasis and from hamsters experimentally infected with 0. viverrini metacercariae. 125I-labelled aqueous somatic extract and in vitro metabolic products of adult worms, and the extracts of metacercariae and eggs were immunoprecipitated, electrophoresed and autoradiographed and major antigens were characterized. Although components with Mr of > 116, 89, 78 and 20 kD appeared to be specifically associated with the somatic extract of adult fluke, the 89 kD protein was the most prominent component found in the in vitro culture fluid of adult worms and the metacercarial extract. Hamsters, experimentally infected with 0. viverrini, also showed the presence of antibody reactive with the 89 kD component within 15 days of infection. The potential of this antigen in immunodiagnosis is discussed.