Abstract: Objective To investigate the prevalence of HPgV-2 in blood donors, transfusion recipients and hemophilia patients and its impact on blood safety. Methods Serum samples were collected from 1060 healthy blood donors, 1402 HCV-positive and 500 HBV- positive blood donors, 570 transfusion recipients and 248 hemophilia patients for screening anti-HPgV-2 antibodies, HPgV-2 RNA, anti-HCV and HBsAg/HBV-DNA using ELISA and RT-PCR. Phylogenetic analysis of near fulllength genome sequences and NS3 genes of pegiviruses and hepaciviruses were performed using MEGA software. Results Anti-HPgV-2 positivity and HPgV-2 RNA positivity were found in 1.21% (17/1402) and 0.36% (5/1402) of the blood donors infected with HCV (RNA+/Ab+), respectively, indicating a close correlation between HPgV-2 and HCV infection (χ2=13.78, P= 0.004). Anti-HPgV-2 antibody was hardly detected in the other populations. A nucleotide identity as high as 97.11% was found in the NS3 fragments among the 5 isolated HPgV-2 strains, which had a nucleotide identity of 96.53% with the reported strains isolated out of China. Conclusion The prevalence of HPgV-2 infection is rather low in healthy blood donors and transfusion recipients. Coinfection with HCV is common in HPgV-2 infection, and no evidence has now been available to support HPgV-2 transmission via blood transfusion, indicating that HPgV-2 may not pose a threat to blood safety.