Abstract:
:It remains unclear whether hepatitis B virus (HBV) replicates in extrahepatic tissues, and particularly in peripheral blood mononuclear cells (PBMCs), which may serve as a reservoir for the maintenance of infection. A real-time PCR assay for the quantitation of total and covalently closed circular (ccc) HBV DNA in serum and in PBMCs was developed. This assay was highly sensitive (detection limit: 27 IU/mL), linear over a wide range (9 log10), and was displayed high inter- and intra-assay reproducibility for the quantitation of total DNA. Genotypes A to E were detected and the results were consistent with those obtained with the COBAS Amplicor HBV Monitor Test. The specificity of the methodology was increased by prior treatment with an enzyme that digests relaxed circular DNA, and the elimination of background signals from virus adsorbed to the surface of PBMCs. HBV DNA was detected in the serum and PBMCs of 12 HBsAg-positive patients, with less than 1% in the cccDNA form. In conclusion, the quantitation of total and ccc HBV DNA in PBMCs is potentially useful as a non-invasive marker, and may help to increase our knowledge of the natural history of hepatitis B.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Mazet-Wagner AA,Baclet MC,Loustaud-Ratti V,Denis F,Alain Sdoi
10.1016/j.jviromet.2006.07.019subject
Has Abstractpub_date
2006-12-01 00:00:00pages
70-9issue
1-2eissn
0166-0934issn
1879-0984pii
S0166-0934(06)00275-8journal_volume
138pub_type
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