To evaluate hepatitis C virus (HCV) and hepatitis B virus (HBV) infection in hemodialysis (HD) units, serum samples from 607 HD patients and 150 staff members at 11 HD units in Japan were collected, and were compared with those from 704 ordinary blood donors as a control. Serum samples subjected to a first generation ELISA for antibody to HCV (anti-C100-3) and were tested by ELISA for HB surface antigen (HBs-Ag), antibody to HBs-Ag, and antibody to HB core antigen (anti-HBc) as HBV markers. We also tested for HCV infection with a second generation ELISA (for antibodies to C22-3, C33c, and C100-3) in 120 HD patients and 30 staff members at 2 selected HD units. Of 607 HD patients, 104 (17%) were positive for anti-C100-3 and 221 (36%) for HBV markers, indicating a much higher prevalence of HCV and HBV infection among HD patients than among ordinary blood donors (0.9% and 18%, respectively). Of 159 patients without a history of blood transfusion, 17 (11%) were positive for anti-C100-3, showing that HCV infection can be acquired without transfusion. The incidence of anti-C100-3 varied from 0% to 53% at different HD units, and HBV markers varied from 17% to 50%. Our study detected a high prevalence of co-infection with HBV and HCV, suggesting that HCV infection may contribute to chronic liver dysfunction in HD patients. Out of 150 staff members, 3 (2%) were positive for anti-C100-3, whereas 25 (17%) were positive for anti-HBc (indicating prior HBV infection).(ABSTRACT TRUNCATED AT 250 WORDS)