The aim of this study was to evaluate the prevalence of antibodies to hepatitis C virus (anti-HCV) in health-care workers (HCW). Sera from 439 unselected HCW were assessed for anti-HCV by 2nd generation enzyme-linked immunoassay (ELISA) and anti-HBc by ELISA. Anti-HCV (+) sera were evaluated by line immunoassay (LIA) (LiaTeK, Organon). Anti-HCV proved positive by ELISA in 12 (2.73%) subjects, 6 of whom were reactive by LIA, one was indeterminate and 5 non reactive. The prevalence of anti-HCV confirmed by LIA was 1.59% (7 subjects). Positive anti-HCV results with an ELISA ratio greater than 3 were LIA reactive in 6/6 as compared with 5 LIA non reactive with an ELISA ratio less than 2, while in the indeterminate serum the ratio was 2.5. No differences in age, profession, seniority, history of hepatitis or transfusions were found between anti-HCV (+) and (-) subjects, but females predominate significantly. The areas of higher risk were hemodialysis, obstetrics, surgery and intensive care. Anti-HBc was (+) in 85.7% (6/7) of the anti-HCV (+) subjects. Follow-up of anti-HCV (+) subjects showed raised alaninoaminotransferase levels in 4 cases, while liver biopsies in 3 disclosed cirrhosis, chronic active hepatitis and chronic persistent hepatitis. The anti-HCV prevalence in HCW is low compared with other risk groups perhaps due to the peculiar epidemiological features of HCV. In low risk groups for HCV infection a positive ELISA result with a ratio lower than 3 should be confirmed by more specific tests.