Although the World Health Organization (WHO) classifies Turkey as a country with a moderate-high prevalence of hepatitis B virus (HBV) infection, there is little data on HBV transmission in this country. To identify risk factors for HBV infection, we performed a retrospective case-control study between January 2007 and December 2009. Acute HBV patients and population controls were selected, and data from these groups were analyzed by logistic regression method. The study included 129 patients with acute HBV infection and 219 controls. Hemodialysis (OR:8.2, 95% CI: 4.17-16.61, p < 0.05), having an HBsAg (+) spouse (OR: 4.3, 95% CI:2.17-8.53, p < 0.05), living with an HBsAg (+) parent(s) (OR: 3.25, 95% CI:1.73-6.12, p < 0.05), and being male (OR: 1.34, 95% CI: 0.82-2.21, p < 0.05) were independent risk factors that were potentially associated with HBV infection. More than one-third of female patients had a significantly higher risk (34.5% vs. 13.5%, p < 0.05) of acquiring HBV from their sexual partners. Hemodialysis was the most frequent risk factor (46.9% vs. 20%, x (2) = 10.45, p < 0.05) for patients aged over 31 years, and living with HBsAg (+) parents was a significantly higher risk factor (28.8% vs. 10.2%, x (2) = 6.15, p < 0.05) that is more likely to lead to HBVin patients aged under 30 years. This study suggests that persons in Turkey who undergo hemodialysis are at high risk for acquiring HBV. Having an HBsAg (+) spouse (sexual transmission) or living with HBsAg (+) parents (household transmission) are significant risk factors for HBV transmission. Vaccination appears to be better preventive method against the spread of HBV.