Quantifying anti-HBV effect of targeted ribonuclease by real-time fluorescent PCR

AIM:To quantify the inhibition of HBV replication by targetedribonuclease by using real-time fluorescent PCR.METHODS:Targeted ribonuclease was introduced into 2.2.15cells by liposome-mediated transfection or HIV-TAT mediatedprotein transduction.Forty-eight hours after the transfectionand 24 h after the transduction,supernatants of 2.2.15 cellswere collected and HBV DNA in the supernatants wasquantified by real-time fluorescent PCR with a commercial kit.RESULTS:HBV DNA concentrations in the supernatants of2.2.15 cells transfected or transducted with targetedribonuclease were 4.9±2.4×10~8 copies/L and 8.3±4.0×10~8copies/L,respectively.Compared with controls,transfectionor transduction of targeted ribonuclease reduced HBV DNAconcentration in the supematants of 2.2.15 cells by 90.4%and 90.1%,respectively(P<0.05).CONCLUSION:Targeted ribonuclease can inhibit HBVreplication in 2.2.15 cells.