Hepatitis E virus (HEV) is now considered the most common cause of acute hepatitis worldwide. There are no published data about the prevalence of antibodies to HEV and RNA in donor sera in Estonia, and this precludes planning measures for preventing HEV proliferation through blood transfusion services. Here, were report data from an analysis of 1002 sera on the prevalence of anti-HEV IgG and IgM and the viral RNA. The antibodies were found in 48 donor sera (4.8%); of these, 40 (4%) harbored anti-HEV IgG, 15 (1.5%) contained anti-HEV IgM, and 7 donors had anti-HEV antibodies of both classes simultaneously. HEV RNA was not detected in any blood serum. Statistical associations of infection risk factors (gender, age, travel in the last six months, contact with pigs and/or wild boars in the last six months, consumption of thermally unprocessed/raw pork or boar meat, raw/unfiltered tap water or water from natural sources, unpasteurized farm dairy products, and unwashed berries and/or vegetables) were assessed. None of the listed factors were found to be associated with a higher or lower risk of anti-HEV antibody presence. At the same time, an increasing share of anti-HEV IgG carriers with age was found. The absence of HEV RNA in the analyzed donor plasma samples proves that HEV acute infection prevalence in Estonia does not exceed the average level of European countries. There is no urgent necessity to enter a requirement for a total screening of blood plasma for HEV RNA prevalence in Estonia.