Background Blood transfusion is a recently reported route of hepatitis E virus (HEV) transmission. It is a bigger concern in regions where large‐scale HEV genotype 1 infections occur causing more severe disease. The present study aims to assess the prevalence and rate of HEV infection in the blood donors of Pune, India. Materials and Methods A total of 2447 healthy blood donors were screened for anti‐HEV IgG and IgM antibodies. Anti‐HEV IgM antibody positives were further subjected to alanine aminotransferase measurement, HEV RNA detection, viral load quantification and phylogenetic analysis. Results Anti‐HEV seroprevalence rate was 17.70%, while IgM prevalence rate was 0.20%. An age dependent increase in IgG seropositive rate was observed. Two of five IgM‐positives tested positive for HEV RNA. The viral load ranged from 3.5 × 104 to 4.6 × 105 copies/mL and belonged to HEV genotype 1. Conclusions HEV prevalence rate of 17.70% in the blood donors of Pune, India, a developing country, goes at par with the developed countries. Current data of 0.20% (5 of 2447) blood donors positive for anti‐HEV IgM and two of them being HEV RNA positive suggest a need for consideration of cost‐effective evaluation towards pooled HEV RNA testing in blood banks.

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