[Dynamic analysis of HBV cccDNA in HepG2 cells infected with Ad-1.2 HBV].
OBJECTIVE
o study the replication of hepatitis B virus (HBV) in HepG2 cells infected with Ad-1.2 HBV.


METHODS
HepG2 cells were transfected with adenovirus containing 1.2 copies of HBV DNA. The expression of HBV antigens were detected in the culture medium by means of enzyme-linked immunosorbent assay (ELISA), and the covalently closed circular DNA (cccDNA) in the cells was extracted with plasmid extraction kit and detected by real-time PCR with selective primer after treatment with mung bean nuclease.


RESULTS
HBsAg, HBeAg and HBV cccDNA were all detected in HepG2 cells after tranfection with Ad-1.2 HBV. HBV cccDNA was detected 1 day after the infection, reaching the peak level 4 days after infection.


CONCLUSIONS
Ad-1.2 HBV-infected cells can serve as the model for screening and evaluation of antiviral agents.
Liang
Wei-fang
WF
Department of Infectious Diseases, Nanfang Hospital, Southern medical University, Guangzhou 510515, China.
Liu
Zhi-hua
ZH
Yang
Jie
J
Luo
Kang-xian
KX
D000256:Adenoviridae / Q000235:genetics*; D002138:Calibration; D045744:Cell Line, Tumor; D018076:DNA, Complementary / Q000235:genetics* / Q000378:metabolism; D004279:DNA, Viral / Q000235:genetics* / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000378:metabolism; D006513:Hepatitis B e Antigens / Q000378:metabolism; D006515:Hepatitis B virus / Q000235:genetics* / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D016133:Polymerase Chain Reaction; D013997:Time Factors; D014162:Transfection; D014779:Virus Replication
D018076:DNA, Complementary; D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens