BACKGROUND: Hepatitis B virus (HBV) and hepatitis C virus (HBV) infections accounts for substantial proportions of the world wide liver disease. The two hepatotropic virus share common modes of transmission and their co-infection are common. Patients with dual HBV and HCV infection have more severe liver disease and are at increased risk of progression to hepatic cell carcinoma. This study was aimed to assess the prevalence of HBV and HCV among patients attending serology laboratory of Gondar University Teaching Hospital. METHODS: A retrospective study was conducted from January 2007 to December 2011 at serology laboratory of Gondar University Teaching Hospital. Data were cross checked for completeness, entered and analyzed using SPSS version 16 statistical package. RESULTS: From the total of 2,684 clinicaly suspected hepatitis patients, 563 (20.98%) were positive for HBsAg and anti-HCV test. Of all, 14.4% were seropositive for HBV (male= 7.89% female=6.27%) (p-value=0.011) while 12.4% were seropositive for HCV (male =7.6% and female=4.13%) (p-value<0.001). The co-incidence of HBV and HCV were found to be 36 (6.39%) (male=3.9% and female=2.4%) (p-value <0.001). Majority of HBV (30.2%) and HCV (30.7%) infections were detected among age group of 25-34 years old (p-value=0.36) and 15-24 years old (p-value<0.001) respectively. CONCLUSION: The overall prevalence of HBV and HCV is high. Males and younger age groups were more affected. Community awareness about the transmission and prevention of viral hepatitis infection should be strengthened by giving health education and herd immunization.