Objective: Through a retrospective analysis of serology and nucleic acid screening data of unpaid blood donors in south Zhejiang region, the role of nucleic acid detection in the reduction of transfusion-related infectious diseases was discussed. Methods: 179 369 unpaid blood donation in south Zhejiong province from Jan, 2016 to Dec, 2016 was chosen. Enzyme linked immunosorbent (ELISA) test for blood index of infectious hepatitis B virus surface antigen (HBsAg), hepatitis C virus antibody (anti-HCV), human immunodeficiency virus antigen and antibody. At the same time the roche, haoyuan nucleic acid detection system were used for HBV-DNA, HCV-RNA, HIV-RNA in 6 or 8 doses mix samples three projects joint detection.NAT results of the serum with negative results in the serological tests were made statisticala analysis. Results: A total of 259 ELISA-/NAT+ samples were detected, HBV-DNA+ 255 cases, HCV-RNA+ 5 cases, one case of HBV-DNA and HCV-RNA + , with a positive rate of 0.14%. The analysis system of roche nucleic acid mix inspection rate of positive was 1.40%, and the split inspection rate of positive was 60.72%. The mix inspection positive rate of the haoyuan analysis system was 1.63%, and the split inspection rate of positive was 41.67%. Conclusions The detection of nucleic acid can make up the deficiency of serological test, and effectively reduce the leakage of transfusion-related infectious diseases, and ensuring the blood safety in this area.