Abstract: Objective To analyze the efficiency of nucleic acid testing(NAT)in blood screening of blood donors in areas of Ji'nan City in Shandong Province. Methods Enzyme-linked immunosorbent assay(ELISA)was used to detect blood samples of unpaid blood donors from 2015 to 2017 in areas of Jinan City. Nucleic acid detection was carried out for the single reagent reactive and double reagent non-reactive samples, and the nucleic acid detection results of non-reactive samples with detections of hepatitis B surface antigen(HBsAg), anti-hepatitis C virus(anti-HCV), anti-human immunodeficiency virus/human immunodeficiency virus(anti-HIV/HIV)by ELISA were analyzed. Results From 2015 to 2017, there were 282 456 screened blood samples of blood donors in Blood Center of Shandong Province, and 1 638 reactive samples(HBsAg, anti-HCV, anti-HIV/HIV)were detected by ELISA, with a total response rate of 0.58%. Totally 105 NAT reactive samples were detected from 280 818 non-reactive samples with detections of three ELISA items(HBsAg, anti-HCV, anti-HIV/HIV), and the total response rate of NAT results was 0.37‰. Conclusion The risk of diseases transmitted by blood transfusion still exists in the condition of using non-reactive blood samples detected by ELISA, and application of NAT detection can shorten the "window period" of ELISA detection and improve the safety of blood samples.