title: Quantification of intrahepatic total HBV DNA in liver biopsies of HBV-infected patients by a modified version of COBAS®Ampliprep/COBAS®TaqMan HBV test v2.0.
abstract: Intrahepatic total HBV DNA (it-HBV DNA) level might reflect the size of virus reservoir and correlate with the histological status of the liver. To quantitate it-HBV DNA in a series of 70 liver biopsies obtained from hepatitis B chronic patients, a modified version of the COBAS®Ampliprep/COBAS®TaqMan HBV test v2.0 was used for this purpose. The linearity and reproducibility of the modified protocol was tested by quantifying serial dilutions of a full-length HBV containing plasmid and it-HBV DNA from a reference patient. A good linear trend between the expected values and those generated by the assay was observed at different concentrations of both plasmid and reference patient (R2= 0.994 and 0.962, respectively). Differences between the values obtained in two independent runs were ≤0.3 log IU for the plasmid and ≤0.6 log IU/mg for the reference patient, showing a high inter-run reproducibility. In the 70 liver biopsies, it-HBV DNA level ranged from 1.4 to 5.4 log IU/mg, with a good linearity and reproducibility between the values obtained in two runs [R2= 0.981; median (IQR) difference of it-HBV DNA 0.05 (0.02-0.09) IU/mg]. The modified COBAS®Ampliprep/COBAS®TaqMan HBV test v2.0 allows an accurate quantitation of it-HBV DNA. Its determination may have prognostic value and may be a useful tool for the new therapeutic strategies aimed at eradicating the HBV infection.
authors.affiliation: Department of Experimental Medicine and Surgery, University of Rome Tor Vergata, Via Montpellier 1, 00133, Rome, Italy.
authors.forename: Romina
authors.initials: R
authors.lastname: Salpini
authors.affiliation: Department of Experimental Medicine and Surgery, University of Rome Tor Vergata, Via Montpellier 1, 00133, Rome, Italy.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Piermatteo
authors.affiliation: Hepatology, Centre for Immunobiology, Blizard Institute, Barts and The London SMD, QMUL, London, UK.
authors.forename: Upkar
authors.initials: U
authors.lastname: Gill
authors.affiliation: Department of Experimental Medicine and Surgery, University of Rome Tor Vergata, Via Montpellier 1, 00133, Rome, Italy.
authors.forename: Arianna
authors.initials: A
authors.lastname: Battisti
authors.affiliation: Laboratory of Molecular Virology, Polyclinic Tor Vergata Foundation, Viale Oxford 81, 00133, Rome, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Stazi
authors.affiliation: Laboratory of Molecular Virology, Polyclinic Tor Vergata Foundation, Viale Oxford 81, 00133, Rome, Italy.
authors.forename: Tania
authors.initials: T
authors.lastname: Guenci
authors.affiliation: Laboratory of Molecular Virology, Polyclinic Tor Vergata Foundation, Viale Oxford 81, 00133, Rome, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Giannella
authors.affiliation: Laboratory of Molecular Virology, Polyclinic Tor Vergata Foundation, Viale Oxford 81, 00133, Rome, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Serafini
authors.affiliation: Hepatology, Centre for Immunobiology, Blizard Institute, Barts and The London SMD, QMUL, London, UK.
authors.forename: Patrick T F
authors.initials: PTF
authors.lastname: Kennedy
authors.affiliation: Department of Experimental Medicine and Surgery, University of Rome Tor Vergata, Via Montpellier 1, 00133, Rome, Italy.|Laboratory of Molecular Virology, Polyclinic Tor Vergata Foundation, Viale Oxford 81, 00133, Rome, Italy.
authors.forename: Carlo Federico
authors.initials: CF
authors.lastname: Perno
authors.affiliation: Department of Experimental Medicine and Surgery, University of Rome Tor Vergata, Via Montpellier 1, 00133, Rome, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Svicher
authors.affiliation: Laboratory of Molecular Virology, Polyclinic Tor Vergata Foundation, Viale Oxford 81, 00133, Rome, Italy. marco.ciotti@ptvonline.it.
authors.forename: Marco
authors.initials: M
authors.lastname: Ciotti
mesh_terms: D000328:Adult; D001706:Biopsy; D004279:DNA, Viral / Q000032:analysis*; D005260:Female; D006515:Hepatitis B virus / Q000235:genetics / Q000302:isolation & purification*; D019694:Hepatitis B, Chronic / Q000821:virology*; D006801:Humans; D008099:Liver / Q000821:virology*; D008297:Male; D015203:Reproducibility of Results; D019562:Viral Load / Q000379:methods*
chemical_list: D004279:DNA, Viral
keywords: COBAS® TaqMan HBV test, v2.0; HBV DNA level; HBV reservoir; Intrahepatic HBV DNA