BackgroundHepatitis is an inflammation of the liver and often caused by viruses. Hepatitis viruses are the leading causes of liver-related morbidity and mortality worldwide, with Hepatitis B and C viruses share the great majority. Studies have shown that prison settings are one of the high-risk environments for the transmission of these viruses. However, there is limited information on the seroprevalence and associated factors of hepatitis B and C viral infection among Ethiopian prisoners.MethodsA facility-based cross-sectional study was conducted among 339 prisoners in Dessie town, Ethiopia from February to April 2020. Hepatitis B surface antigen and antibody against hepatitis C virus in serum were determined using Enzyme-Linked Immunosorbent Assay. We imputed the data using “EpiData 3.1” software and exported it to Statistical Package for Social Sciences version 20.0 for analysis, and a p-value of <0.05 was considered statistically significant.ResultsThe overall seroprevalence of hepatitis B surface antigen and anti-hepatitis C virus among prisoners was 22/339 (6.5%) (95% CI = 3.8–9.4), and 4/339 (1.2%) (95% CI = 0.0–2.4), respectively. Multiple sexual partners, previous imprisonment, body tattooing, and contact with the jaundiced patient were independently associated with hepatitis B virus infection. Prisoners who had a history of blood transfusion, and dental extraction were independently associated with hepatitis C virus infection.ConclusionThe seroprevalence of hepatitis B and hepatitis C viral infection among Dessie town prisoners was intermediate and low, respectively. The finding of a significant association between the presence of Hepatitis B surface antigen and hepatitis C virus antibodies among prisoners and factors calls for the need of serological testing for both Hepatitis B and C viruses to high-risk individuals. Strengthening screening strategies and prevention programs in prison settings is advisable to prevent disease transmission.