In Chile, there are few studies about seroprevalence of IgG antibodies against hepatitis E virus (HEV) in blood banks, between 4 and 8%. The development of new techniques with greater sensitivity and specificity, account for an increase in the seroprevalence of HEV in various countries, the current status in Chile being unknown. In the present study, we determined the seroprevalence of anti-HEV IgG in blood donors of the Clinical Hospital University of Chile, with last generation ELISA techniques. Out of a total of 186 samples, collected in 2014, 56 (30.1%) were positive, without gender differences, but with a significant increase with age (p < 0.001). These results show an increase in the seroprevalence of HEV in blood donors performed with immunoassays of greater sensitivity.