Hepatitis A virus (HAV) infection is one of the important causes of hepatitis in developing countries. It is transmitted through feco-oral route. This poses major health problems in children. Serum samples of the patients from pediatrics department suspected of acute viral hepatitis presenting with clinical features like fever, nausea, vomiting, and jaundice were collected and analyzed by using commercially available mini VIDAS 307 (Biomeriux) to detect anti HAV IgM antibodies. A nine months prospective study was conducted in the Department of Microbiology and immunology of Shri Guru Ram Rai Institute of Medical and health science Dehradun, India. Of the 47 serum samples received during the study period, 23 (48.93%) were found to be positive for anti-HAV IgM (VIDAS 307 Biomeriux). The prevalence was found to be more in male patients than in female patients, that is,14 (60.8%) and 9 (39.13%) respectively, and was predominantly seen in the 5–10 years of the age group (60%). Peak cases were found in the month of October and November. The study showed a high positivity (48.93%) of HAV in this sub-Himalayan region indicating the role of preventive measures including safe drinking water, improved sanitation and vaccination in preventing infection. Thus, detection of anti-HAV IgM becomes the cornerstone entity for diagnosis.