Abstract: The aim of this study was to determine the seroprevalence of HCV in HBsAg positive subjects. In a population based single stage cluster sampling in Golestan province of Iran, 139 HBsAg positive subjects were detected and enrolled in the study. Serum samples were tested using ELISA method for anti-HCV antibodies. Chi-square and Fisher`s exact tests were used to compare the proportions. There were 68(48.9%) male and 71(51.1%) female. The mean age was 41.89±11.30 years (25-64 years). One case was excluded because of inadequate blood sampling. Anti-HCV antibody was positive in 17(12.3%) of 138 remaining subjects. Seropositivity of HCV in female was higher than male (14.1% versus 10.4%, respectively) but the difference was not statistically significant (p = 0.52). No significant relationship was seen between Anti-HCV seroprevalence and demographic factors such as age, place of residence and marital status. The seroprevalence of co-infection with HBV and HCV in our study was higher than such reports from some countries (Italy); but it was in line with worldwide prevalence (>10%). A careful longitudinal evaluation of the viremia levels of both HBV/HCV viruses is essential for making a correct diagnosis and tailoring the appropriate therapeutic schedule in co-infected patients.