Hepatitis E virus (HEV) is a well-known cause of sporadic acute hepatitis. The contribution of fecal shedding of the virus to its endemic nature is not frequently studied in underdeveloped countries. The aim of the present study was to detect HEV viremia in serum and stool from patients with acute hepatitis by cell culture and by nested reverse transcriptase (RT)-PCR. A further aim was to evaluate different methods used for HEV detection, including culture by use of HPG11 cell line, PCR, immunoglobulin M (IgM) and IgG responses during the acute stage of infection. The frequency of HEV-positive cases for IgM, stool and serum cultures was 35.3%, 38.2% and 29.4%, respectively. However, only two samples (2.9%) were positive for IgG using enzyme immunoassay. The sensitivity of stool culture was 41.9%, the sensitivity of both HEV IgM and the combined laboratory tests was 37.5% for each, the sensitivity of serum culture was 30.3% and the sensitivity for HEV IgG was 2.3%. We conclude that hepatitis E viremia can be detected both in serum and in stool from patients with acute hepatitis. The shedding of virus in the stool of patients may be the responsible factor for the endemicity of HEV. In addition, the detection of HEV had high sensitivity compared with other methods. Nevertheless, it was necessary to use both stool and serum cultures to avoid missing any case with HEV infection.