[Quantitative detection of hepatitis B virus covalently closed circular DNA in sera of chronic hepatitis B patients with a newly established assay].
OBJECTIVE
To quantitatively detect hepatitis B virus covalently closed circular DNA (HBV cccDNA) in sera of chronic hepatitis B patients with a newly established assay.


METHODS
Primers and probe were designed in highly conservative region of HBV DNA. DNA was extracted from 175 sera samples of chronic hepatitis B patients, and was treated with plasmid-Safe-ATP-dependent Dnase(PSAD) to eliminate the relaxed circular DNA (rcDNA). The products were amplified by real-time PCR with primers spanning.


RESULTS
The detection rate of serum HBV cccDNA was found to correlate directly with serum HBV DNA loading. HBeAg positive chronic hepatitis B patients had higher serum HBV cccDNA levels than HBeAg negative chronic hepatitis B patients.


CONCLUSIONS
The method is good because of the high specificity. It can be used for detection of HBV cccDNA. DNA;
Zhong
Yan-Wei
YW
Viral Hepatitis Research Laboratary, Institute of Infectious Disease of PL4 302 Hospital, Beijing 100039, China.
Liang
Zhao-Ling
ZL
Ren
Xiao-Qiang
XQ
Li
Xiao-Dong
XD
Xu
Zhi-Hui
ZH
Li
Le
L
Xu
Dong-Ping
DP
D000293:Adolescent; D000328:Adult; D017931:DNA Primers / Q000235:genetics; D004270:DNA, Circular / Q000097:blood* / Q000235:genetics; D004279:DNA, Viral / Q000097:blood* / Q000235:genetics; D005260:Female; D006515:Hepatitis B virus / Q000235:genetics*; D019694:Hepatitis B, Chronic / Q000821:virology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D055815:Young Adult
D017931:DNA Primers; D004270:DNA, Circular; D004279:DNA, Viral