Quantifying anti-HBV effect of targeted ribonuclease by real-time fluorescent PCR.
OBJECTIVE
To quantify the inhibition of HBV replication by targeted ribonuclease by using real-time fluorescent PCR.


METHODS
Targeted ribonuclease was introduced into 2.2.15 cells by liposome-mediated transfection or HIV-TAT mediated protein transduction. Forty-eight hours after the transfection and 24 h after the transduction, supernatants of 2.2.15 cells were collected and HBV DNA in the supernatants was quantified by real-time fluorescent PCR with a commercial kit.


RESULTS
HBV DNA concentrations in the supernatants of 2.2.15 cells transfected or transducted with targeted ribonuclease were 4.9+/-2.4 x 10(8) copies/L and 8.3+/-4.0 x 10(8) copies/L, respectively. Compared with controls, transfection or transduction of targeted ribonuclease reduced HBV DNA concentration in the supernatants of 2.2.15 cells by 90.4% and 90.1%, respectively (P<0.05).


CONCLUSIONS
Targeted ribonuclease can inhibit HBV replication in 2.2.15 cells.
Liu
Jun
J
Department of Etiology, Fourth Military Medical University, Xi'an 710033, Shaanxi Province, China. etiology@fmmu.edu.cn
Li
Ying-Hui
YH
Ding
Jin
J
Gong
Wei-Dong
WD
Xue
Cai-Fang
CF
Zhao
Ya
Y
Huang
Yu-Xiao
YX
D045744:Cell Line, Tumor; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D006515:Hepatitis B virus / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D018197:Hepatoblastoma; D006801:Humans; D008113:Liver Neoplasms; D016133:Polymerase Chain Reaction / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D012260:Ribonucleases / Q000008:administration & dosage / Q000494:pharmacology*; D014162:Transfection; D014779:Virus Replication / Q000187:drug effects*
D017931:DNA Primers; D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic; D012260:Ribonucleases