Objective To investigate the relationship between HBV-DNA with pre S1 antigen in patients with HBsAg，Anti-HBe and Anti-HBc positive hepatitis B. Methods The serum express of pre S1 antigen and HBV- DNA in 152 patients with HBsAg，Anti-HBe and Anti-HBc positive hepatitis B were detected by ELISA and PCR(FQ-PCR) respectively. Results 74 cases of samples were both pre S1 and HBV-DNA positive，while 34 cases were found Pre S1 and HBV-DNA negative. The positive coincidence rate between Pre S1 and HBV-DNA was 71. 1%. There were 30 cases with pre S1 positive and HBV-DNA negative，while 14 cases had HBV-DNA positive and pre S1 negative. The detection rate of pre S1 and HBV-DNA in patients with HBsAg，Anti-HBe and Anti-HBc positive hepatitis B were 68. 4% and 57. 9%，respectively. Conclusion There are virus replications of different degrees in patients with HBsAg，Anti-HBe and Anti-HBc positive hepatitis B. The detection of Pre S1 can not replace HBV-DNA detection，and it could be only used as a supplementary index. The HBV- DNA quantitative detection could facilitate to judge whether virus in-vivo replicate or not.