Hepatitis E virus (HEV), transmitted through the fecal–oral route is a common cause of self-limiting acute viral hepatitis in India with poor sanitation and hygiene and poses a major public health problem in India. This study was undertaken to determine seroprevalence of HEV in patients presenting with acute viral hepatitis. A retrospective study was conducted in Central Laboratory, General Hospital, Palanpur, North Gujarat from October 2016 to July 2018. A total of 164 serum samples of suspected patients of HEV were tested for IgM HEV antibody using ELISA method. Seroprevalence of HEV was 62.20%. Out of 102, 70 (68.63%) males were affected more than females (31.37%). HEV infection was more common in 16-25 years age group (38.24%) followed by 26-35 years age group(24.51%). To reduce the prevalence of HEV infection, health and civic authorities should make an effort to increase the awareness among general public, to facilitate the timely preparation and make necessary arrangement for efficient response to encounter outbreak or epidemic caused by HEV virus, thus reduce morbidity, mortality and economic burden.