Acute viral hepatitis(AVH) is caused by Hepatitis A (HAV) and Hepatitis E(HEV). It is major health burden in India. Both the viruses HAV and HEV are primarily transmitted via the faeco-oral course. Study was conducted to determine the seroprevalence of HAV, HEV and rate of co-infection in AVH patients attending rural tertiary care centre. A retrospective laboratory record based study was carried out in rural tertiary health care center located in Western Maharashtra. Laboratory and Medical records of suspected acute viral infection patients were analyzed during study. Study period was June 2014 to July 2018. Commercially available ELISA kits of IgM anti-HAV and IgM anti-HEV were used to analyze serum samples of suspected study participants. Tests were carried out as per the manufacturer’s instructions. A total of 778 acute viral hepatitis cases were included in the study from July 2014 to July 2018 among which 85/778 (10.9 %) detected positive for HAV and 121/778 (15.6%) detected positive for HEV. Co-infection was identified in 6/778(0.8 %). Jaundice, fever fatigue and hepatomegaly were common clinical presentation in HAV, HEV and confection with both viruses in acute viral hepatitis patients. Study indicated low exposure to HAV in childhood bellow 16 years. Co-infection rate was detected high in 16-25 years age group. Vaccination policy against HAV in adolescent age group needed as there is change epidemiological shift of HAV which has been observed in the current study. These data will helps for planning future vaccination strategies, better implementation sanitation program, and safe water supply in this geographic area.