Transmission of West Nile virus (WNV) from asymptomatic donors has been reported during blood transfusions and organ transplants in humans. In this work, we aimed to investigate the presence of WNV antibody and WNV RNA in blood donors to evaluate the sero-prevalence of WNV and risk for WNV transmission. One hundred and sixty blood donors were tested for the presence of anti-WNV IgG by ELISA and for WNVs 1 and 2 RNA by RT-PCR. About 55% of blood donors were seropositive for WNV IgG antibodies, with significantly higher percentage of positive donors coming from rural areas and Nile Delta region compared to other donors. Using RT-PCR all donors were negative for viral RNA of both WNV lineages 1 and 2. High sero-prevelance of WNV antibodies in asymptomatic blood donors denotes endemicity of the WNV in Egypt and points to the importance of routine screening of blood donors for WNV RNA. On the other hand the absence of WNV RNA by RT-PCR indicates apparent low risk of the blood products as regards WNV transmission. Further studies into significance of WNV seronegativity among Rh negative donors and into the use of WNV seropositive blood in prophylaxis or treatment of WNV neuroinvasive disease are recommended. J. Med. Virol. 89:1323-1329, 2017. © 2017 Wiley Periodicals, Inc.