Hepatitis A virus (HAV) infection is one of the important causes of hepatitis in developing countries. It is transmitted through feco-oral route. They pose major health problems. To determine the prevalence of HAV in patients presenting with acute viral hepatitis. Serum samples of the patients presenting with fever, nausea, vomiting, and jaundice were collected and analyzed by using commercially available RecombiLISA ELISA kit for the detection of immunoglobulin M (IgM) against HAV. A 20 months retrospective study was conducted in the Department of Microbiology, Shri M. P. Shah Govt. Medical College, Jamnagar, Gujarat (India). Of the 532 serum samples subjected to RecombiLISA ELISA for detection of anti-HAV IgM in patients, 157 (29.51%) were found to be positive. The prevalence was found to be more in male patients than in female patients, that is, 81(51.59%) and 76(48.41%), respectively, and was predominantly seen in the 6–15 years of the school-going age group. Peak cases were found in the monsoon season. Regular monitoring of clinical, serological and molecular characteristics would help in understanding the epidemiology of HAV and in planning the intervention studies.