Transmission of hepatitis E virus (HEV) through transfusion of blood components has already been reported in several European countries. Here, we assessed the HEV prevalence in Flemish blood donors. This study is of importance in order to assess the risk of HEV transmission through blood transfusion. We analyzed 38,137 blood donation samples that were collected by the Red Cross Flanders during the period May-June 2015. All samples were screened for the presence of HEV RNA and a selection for HEV-specific IgM/IgG. After pooling per 6, 11 pools reacted positive during RNA screening. Reactive pools were deconstructed and individual samples were retested. After deconstruction, 7 samples were confirmed as HEV RNA positive. Serological screening of the confirmed RNA positive samples showed that 6 out of these 7 samples were HEV IgM positive, of which 3 donors were also IgG positive. Serological screening was also performed on the samples that constituted the 4 initially HEV RNA reactive pools where RNA positivity was not confirmed on the individual level. In 3 pools we found indirect evidence of recent HEV exposure. Within 356 randomly selected samples, 31 donations were HEV IgG positive. Here we show that at least 1:5.448 of blood donations in Flanders may originate from donors that are actively infected with HEV. Upon transfusion, these donations may pose a major threat towards patients at risk. Finally, a serological analysis showed that the anti-HEV IgG prevalence in Flemish blood donors is 8,71%. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.