查了一下，有一篇文章是做这个的，就是用PCR检测2.2.15的cccDNA
Liu, M.C., et al. , Dynamic analysis of hepatitis B virus DNA and its antigens in 2.2.15 cells. Journal of Viral Hepatitis, 2004. 11(2): p. 124--129.
SUMMARY. The 2.2.15 cells-derived from HepG2 cells
transfected with a plasmid containing hepatitis B virus
(HBV) DNA secrete surface antigen (HBsAg) particles, nucleocapsids
and virions (Proc Natl Acad Sci U S A 1987; 84:
1005–1009). The latter elicit acute hepatitis in chimpanzees
(Proc Natl Acad Sci U S A 1987; 84: 4641–4644). We studied
the presence of intracellular and extracellular HBV covalently
closed circular (ccc) DNA in this culture system by
polymerase chain reaction (PCR), kinetically analysed
HBsAg and hepatitis B e antigen (HBeAg) released in the
culture media by quantitative enzyme-linked immunosorbent
assay and quantitated by real-time PCR but HBV DNA
from intracellular and extracellular HBV-DNA. HBV cccDNA
was found both intracellularly and extracellularly. A significant
correlation was seen between the extracellular HBV
DNA levels and virus antigens (r ¼ 0.833; P ¼ 0.01 and
r ¼ 0.939; P < 0.01 for HBsAg and HBeAg, respectively),
whereas there was no statistical correlation between intracellular
HBV DNA levels and virus antigen levels (r ¼ 0.024;
P ¼ 0.955 and r ¼ 0.177; P ¼ 0.625 for HBsAg and
HBeAg, respectively). These data would be valuable in
studies of the HBV life cycle and of potential anti-viral
agents. 作者: cao1976 时间: 2015-8-23 18:38
bigen发表于 2009-7-22 20:24：
这篇文章里面说cccDNA的提取方法：
To enrich for ccc forms, DNA was isolated from cytoplasmic and nuclear fractions of
2.2.15 cells by two extractions with phenol, followed by two extractions with phenol and chloroform (1:1) without prior treatment with proteinase K.
Sells, M.A., et al. , Replicative intermediates of hepatitis B virus in HepG2 cells that produce infectious virions. Journal of Virology, 1988. 62(8): p. 2836--2844. 作者: cao1976 时间: 2015-8-23 18:38
gong发表于 2009-7-23 19:05 ：