Objectives The aim of this study was to evaluate the seroprevalence of hepatitis E virus (HEV), a major public health issue worldwide with the potential for transmission via blood transfusion, in blood donors in the Qassim Region, Saudi Arabia. Serum samples (n = 1,078) were collected from volunteer blood donors from January to April 2019 and tested for the presence of anti-HEV IgG and IgM by indirect enzyme-linked immunosorbent assays. Results Overall, the seroprevalence of anti-HEV IgG and IgM among blood donors was 5.7% and 1.3%, respectively. Additionally, the seropositive rates of anti-HEV IgG and IgM were significantly higher in non-Saudi donors (22.1% and 7.8%) than in Saudi donors (3% and 0.2%). The seroprevalence of anti-HEV IgG increased with age; however, there was no correlation between gender and anti-HEV IgG and/or IgM. The seroprevalence of HEV among blood donors in the Qassim Region was lower than previous estimates for other regions of the country. Further studies covering a wider geographical area are needed to validate and expand the findings and to determine the importance of HEV screening in the region.