通过修改版本的COBAS®Ampliprep /COBAS®TaqManHBV测试v2.0，对HBV感染

StephenW

Med Microbiol Immunol. 2017 Apr 11. doi: 10.1007/s00430-017-0504-3. [Epub ahead of print]
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.
Salpini R1, Piermatteo L1, Gill U2, Battisti A1, Stazi F3, Guenci T3, Giannella S3, Serafini V3, Kennedy PT2, Perno CF1,3, Svicher V1, Ciotti M4.
Author information

1    Department of Experimental Medicine and Surgery, University of Rome Tor Vergata, Via Montpellier 1, 00133, Rome, Italy.
2    Hepatology, Centre for Immunobiology, Blizard Institute, Barts and The London SMD, QMUL, London, UK.
3    Laboratory of Molecular Virology, Polyclinic Tor Vergata Foundation, Viale Oxford 81, 00133, Rome, Italy.
4    Laboratory of Molecular Virology, Polyclinic Tor Vergata Foundation, Viale Oxford 81, 00133, Rome, Italy. [email protected].

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 (R 2 = 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 [R 2 = 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.
KEYWORDS:

COBAS® TaqMan HBV test, v2.0; HBV DNA level; HBV reservoir; Intrahepatic HBV DNA

PMID:
    28401351
DOI:
    10.1007/s00430-017-0504-3

StephenW

Med Microbiol Immunol。 2017年4月11日。doi：10.1007 / s00430-017-0504-3。 [提前印刷]
通过修改版本的COBAS®Ampliprep /COBAS®TaqManHBV测试v2.0，对HBV感染患者的肝脏活检中肝内总HBV DNA进行定量。
Salpini R1，Piermatteo L1，Gill U2，Battisti A1，Stazi F3，Guenci T3，Giannella S3，Serafini V3，Kennedy PT2，Perno CF1,3，Svicher V1，Ciotti M4。
作者信息

1
    罗马大学实验医学与外科学系Tor Vergata，Via Montpellier 1，00133，Rome，Italy。
2
    肝病学，免疫生物学中心，Blizard研究所，Barts和伦敦SMD，QMUL，伦敦，英国。
3
    分子病毒学实验室，Polyclinic Tor Vergata Foundation，Viale Oxford 81，00133，Rome，Italy。
4
    分子病毒学实验室，Polyclinic Tor Vergata Foundation，Viale Oxford 81，00133，Rome，Italy。 [email protected]

抽象

肝内总HBV DNA（it-HBV DNA）水平可能反映病毒库的大小，并与肝组织学状态相关。为了定量 - HBV DNA在一系列从乙型肝炎慢性患者获得的70例肝脏活检中，使用了修改版本的COBAS®Ampliprep/COBAS®TaqManHBV测试v2.0。通过定量来自参考患者的全长HBV含有质粒和其-DNA DNA的连续稀释度来测试修饰方案的线性和重复性。在质粒和参考患者的不同浓度下观察到预期值与测定产生的线性趋势之间良好的线性趋势（分别为R 2 = 0.994和0.962）。在两个独立运行中获得的值之间的差异对于质粒为≤0.3logIU，对于参考患者为≤0.6logIU/ mg，显示出高的体内再现性。在70例肝脏活检中，HBV DNA水平范围为1.4〜5.4logIU / mg，两次运行时获得的值之间具有良好的线性和重现性[R 2 = 0.981; HBV DNA的中位数（IQR）差异0.05（0.02-0.09）IU / mg]。经修改的COBAS®Ampliprep/COBAS®TaqManHBV测试v2.0可以准确定量检测HBV DNA。其决定可能具有预后价值，可能是旨在消除HBV感染的新治疗策略的有用工具。
关键词：

COBAS®TaqMan HBV检测v2.0; HBV DNA水平;乙肝病毒肝内HBV DNA

PMID：
    28401351
DOI：
    10.1007 / s00430-017-0504-3