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Context Hepatitis E virus (HEV) as a hepatotropic virus is one of the major global health concerns. Autochthonous HEV transmitted by oral fecal-route in poor sanitation conditions as well as vertical and rarely blood transfusion. HEV occurrence is more common in developing countries and recently increased in developed countries too. Middle East (ME) and Eastern Mediterranean region (EMR) of WHO have been an endemic region for HEV infection. In this regard, we aimed to design a systematic review and pooled analysis to determine seroprevalence of anti-HEV antibody in ME and EMR countries. Evidence acquisition By using PRISMA guideline, data were collected from papers identified through PubMed, Web of Science, Science Direct, Scopus and also from some national and regional databases from January 1990 to June 2016. Serum anti-HEV antibody (IgG) used for HEV prevalence estimation. HEV prevalence in the ME, WHO EMR countries, and in total, calculated by each country population size based on 2015 UN report. Results overall, 62 papers with a total sample size of 31,673 were fulfilled our eligibility criteria and included in our project. Considering anti-HEV antibody (IgG), prevalence of HEV infection in the countries of ME, WHO EMR and in total were 12.17% (95% CI: 11.79–12.57), 11.81% (95% CI: 11.43–12.21), and 11.87% (95% CI: 11.52–12.23) respectively. Conclusions HEV seroprevalence in WHO EMR and ME countries has high rate and more considerations are needed for the prevention and control of this infection especially in high-risk groups such as pregnant women. [ABSTRACT FROM AUTHOR]