【Objective】 To analyze the preliminary screening and follow-up testing data of HBV in Yantai area, and discuss the rationality of following up and re-entry program of HBV reactive blood donors. 【Methods】 Donors who were single reagent reactive by enzyme-linked immunosorbent assay (ELISA) in initial screening but non-reactive by nucleic acid testing (NAT) were followed up. Individual NAT(ID-NAT) was performed for HBV DNA, ELISA for HBsAg, HBsAb, HBeAb, HBeAg and HBcAb, and ECLIA for the detection of HBsAg. 【Results】 A total of 547 blood donors were HBsAg ELISA-/NAT+, and 97 were followed up, among which 24 met the requirements of re-entry while 73 did not. Of the 24 blood donors who met the re-entry requirements, 13 donated blood again, with test results all qualified. 【Conclusion】 The combination of ELISA, ID-NAT, and ECLIA methods for following up detection for HBsAg ELISA+ blood donors is recommended. Blood donors with HbsAb S/CO ≥ 10 and negative results for other tests met the re-entry requirements, with a re-entry rate at 24.74%, and the re-donation qualified rate of blood donors after re-entry was 100%.