ObjectiveTo establish domestic entirely automatic nucleic acid testing (NAT) assay for blood screening, and to study the feasibility of domestic NAT.MethodsOn the basis of enzyme linked immunosorbant assay for blood screen, blood samples were  pooled and extracted nucleic acid with entirely automatic nucleic acid apparators, then the extracted nucleic acid was amplified and detected with realtime polymerase chain reaction，sensitivity and efficacy was evaluated, NAT positive donors were traced.ResultsThe 95% detection limits of HBV DNA, HCV RNA and HIV1 RNA tested by automatic system  were 35.9 IU/mL，147.7 IU/mL and 66.1 IU/mL，respectively,95% confidence intervals were ［21.9，124.8］，［93.3，386.4］ and ［38.8，235.3］ respectively. Twelve of 60 112 samples were HBV DNA positive，DNA positive ratio was 1︰5 009, 6 of which  were antiHBc positive;one sample was HCV RNA positive, RNA positive ratio was 1︰60 112; no HIV1 RNA positive was detected among all samples. Seven HBV DNA donors were followed up, 4 of whom had  seroconverted.ConclusionThe domestic entirely automatic system can be applied in blood HBV DNA , HCV RNA and HIV RNA screening.