Objective To study the relationship between hepatitis B virus serum markers and hepatitis B virus DNA (HBV DNA) with hepatitis B virus Pre-S2 antigen (Pre-S2) .Methods Hepatitis B serum markers and Pre-S2 antigens were tested by enzyme-linked immunosorbent assay and HBV DNA was detected by fluorescent quantitative-PCR in 982 hepatitis B patients.Results The detection rates of Pre-S2 and HBVDNA in HBeAg-positive and negative groups were 95.34% and 97.8%, and 39.7% and 32.3%, respectively.The detection rate of Pre-S2 antigen was 1.45% among HBsAg and HBeAg negative donors.Conclusion The Pre-S2 antigen is closely related with HBeAg and HBVDNA and the Pre-S2 antigen could be the complement to serum markers.