Subcloning of HBV and the Application on Nucleic Acid Hybridization

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.