Although hepatitis B has been well studied, there are still aspects of its epidemiology that remain to be clarified. There are many regions with high seroprevalence, particularly in the developing regions of the world, and these regions are known to have different epidemiologic patterns. Nonetheless, there are currently no data on the differences in hepatitis 13 seroprevalence between urban and rural areas of Turkey. In the present study, therefore, we used 30-cluster sampling to determine and compare the prevalence of hepatitis B in the urban and rural areas of the least developed region of Turkey, the southeastern region. From 2,888 adults living in the region, blood samples were obtained from house visits, and screened for HBsAg, anti-HBs, and anti-HBclgG. Factors associated with hepatitis B seroprevalence, particularly living in rural areas, were analyzed with multivariate methods. The seroprevalence of HBsAg was 8.2% in the rural and 6.26% in the urban areas. There was a statistically significant difference between urban and rural regions in terms of HBsAg positivity (crude OR: 0.74; 95% CI: 0.55 - 0.98). Exposure to hepatitis B virus (HBV) increased with age both in urban and rural areas. Lower education level was also an important risk factor for hepatitis B seropositivity in urban areas (adjusted OR: 1.66; 95% CI: 1.26 - 2.19) but not in rural ones (adjusted OR: 0.77; 95% CI: 0.36 - 1.69). Familial jaundice history was a statistically significant risk factor for HBsAg positivity in rural areas (adjusted OR: 2.15; 95% CI: 1.30 - 3.56) but not in urban ones (adjusted OR: 1.48; 95% CI: 0.96 - 2.27). This study shows that the prevalence of HBV infection-in the southeastern region of Turkey is intermediate among the levels reported for the European region of the World Health Orgnization.