Background Viral hepatitis (hepatitis A, B, C, D and E) remains a public health problem in Peru, with a high disease burden. There are limited data on the prevalence of viral hepatitis at a national level, and none reported for over two decades. In this study, the prevalence rates of hepatitis A (HAV), B (HBV), C (HCV), D (HDV) and E virus (HEV) infections in the Peruvian population were determined to provide updated baseline data that would help guide the development of strategies aimed at reducing the transmission of viral hepatitis in Peru. Methods We conducted a cross-sectional, population-based study in the 25 regions of Peru. The study included participants of both sexes, aged 15-69 years, who had lived for >6 months in a specific region of Peru. Serum samples were analyzed by ELISA for anti-HAV (IgG), anti-HBs �10 mUI/ml, anti-HCV, anti-HDV and anti-HEV (IgG) antibodies, and by chemiluminescence for the HBV surface antigen (HBsAg) and antibodies against the core HBV antigen (anti-HBc IgM and IgG). Results In a total of 5183 study participants, the prevalence rates of anti-HAV (IgG), HBsAg, total anti-HBc IgG, anti-HBs �10 mUI/ml, anti-HCV and anti-HEV (IgG) were 98.4% [95% confidence interval (CI) 98.0-98.7), 0.4% (95% CI 0.21-0.55), 10.1% (95% CI 9.4-11.0), 60% (95% CI 58.5-61.2), 0.1% (95% CI 0.02-0.25), and 14% (95% CI 13.1-15.0%), respectively. The prevalence of anti-HDV among HBsAg carriers was 15% (3/20).