Some studies have suggested that hepatitis E virus is more frequent in human immunodeficiency virus patients and can progress to chronic infection. We aimed to determine the prevalence of hepatitis E virus antibodies and RNA in a series of 100 human immunodeficiency virus patients in Tehran, Iran, with comparison to 52 healthy human immunodeficiency virus and hepatitis B and C negative blood donors as controls. Human immunodeficiency virus patients were also tested hepatitis E virus-RNA. Among the human immunodeficiency virus patients, 10% had antibodies to hepatitis E virus – a finding not significantly different from the uninfected controls (11.5%). No human immunodeficiency virus patients had hepatitis E virus IgM antibodies nor did any have detectable hepatitis E virus-RNA. We found no associations between anti-hepatitis E virus IgG-seropositivity and age, sex, route of human immunodeficiency virus acquisition, aminotransferases levels, CD4, antiretroviral therapy, hepatitis B virus and hepatitis C virus co-infection. Hepatitis E virus is relatively prevalent in our human immunodeficiency virus patients, although without evidence of chronic infection and no more common than among human immunodeficiency virus-negative controls or the general population. For the present, we do not recommend routine screening for hepatitis E virus infection in human immunodeficiency virus patients in our moderately endemic region.