Abstract: Objective Subcloning strain with whole gene of HBV is applied for nucleic acid hybridization diagnosis and the studies of transgenic tomatoes.Methods In this experiment,clone the HBV DNA of pBR₃₂₂-HBV into higher copies vector pUC₁₉ to get the new strain of pUC₁₉-HBV.HBV-DNA probe labeled with Dicogingenin was made by digesting the pUC₁₉-HBV with endonuclease and purificafion of HBV DNA.72 cases of patient serum were paralleled detected by nucleic acid hybridization,QPCR and PCR electrophoresis.gain the subcloning strain of pUC₁₉-HBV,use the DIG-HBV probe labeled with Digoxigenin,QPCR and PCR to detect 72 cases serums simultaneously.Results The positive cases were 40,51,49 respectively.The positive percentage was 55.6%,70.8%,68.1% respectively.Conclusions Among the three methods of detecting HBV,QPCR is the most sensitive method,even though the hybridizations is the most specific.