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使用雅培实时丙型肝炎病毒检测法对肝脏活检样本和肝外腔室中的丙型肝炎病毒核糖核酸进行定量分析。

HCV-RNA quantification in liver bioptic samples and extrahepatic compartments, using the abbott RealTime HCV assay.

作者信息

Antonucci FrancescoPaolo, Cento Valeria, Sorbo Maria Chiara, Manuelli Matteo Ciancio, Lenci Ilaria, Sforza Daniele, Di Carlo Domenico, Milana Martina, Manzia Tommaso Maria, Angelico Mario, Tisone Giuseppe, Perno Carlo Federico, Ceccherini-Silberstein Francesca

机构信息

Department of Experimental Medicine and Surgery, University of Rome Tor Vergata, Rome, Italy.

Liver Unit, Polyclinic Tor Vergata Foundation, University of Rome Tor Vergata, Rome, Italy.

出版信息

J Virol Methods. 2017 Aug;246:1-7. doi: 10.1016/j.jviromet.2017.04.001. Epub 2017 Apr 11.

Abstract

BACKGROUND & AIMS: We evaluated the performance of a rapid method to quantify HCV-RNA in the hepatic and extrahepatic compartments, by using for the first time the Abbott RealTime HCV-assay.

METHODS

Non-tumoral (NT), tumoral (TT) liver samples, lymph nodes and ascitic fluid from patients undergoing orthotopic-liver-transplantation (N=18) or liver resection (N=4) were used for the HCV-RNA quantification; 5/22 patients were tested after or during direct acting antivirals (DAA) treatment. Total RNA and DNA quantification from tissue-biopsies allowed normalization of HCV-RNA concentrations in IU/μg of total RNA and IU/10 liver-cells, respectively.

RESULTS

HCV-RNA was successfully quantified with high reliability in liver biopsies, lymph nodes and ascitic fluid samples. Among the 17 untreated patients, a positive and significant HCV-RNA correlation between serum and NT liver-samples was observed (Pearson: rho=0.544, p=0.024). Three DAA-treated patients were HCV-RNA "undetectable" in serum, but still "detectable" in all tested liver-tissues. Differently, only one DAA-treated patient, tested after sustained-virological-response, showed HCV-RNA "undetectability" in liver-tissue.

CONCLUSIONS

HCV-RNA was successfully quantified with high reliability in liver bioptic samples and extrahepatic compartments, even when HCV-RNA was "undetectable" in serum. Abbott RealTime HCV-assay is a good diagnostic tool for HCV quantification in intra- and extra-hepatic compartments, whenever a bioptic sample is available.

摘要

背景与目的

我们首次使用雅培实时丙型肝炎病毒检测法,评估了一种快速定量肝内和肝外区域丙型肝炎病毒RNA的方法的性能。

方法

对接受原位肝移植(N = 18)或肝切除术(N = 4)患者的非肿瘤性(NT)、肿瘤性(TT)肝脏样本、淋巴结和腹水进行丙型肝炎病毒RNA定量;22例患者中有5例在直接作用抗病毒药物(DAA)治疗期间或之后接受检测。组织活检的总RNA和DNA定量分别使丙型肝炎病毒RNA浓度以每微克总RNA的国际单位数和每10个肝细胞的国际单位数进行标准化。

结果

在肝活检、淋巴结和腹水样本中成功且高度可靠地定量了丙型肝炎病毒RNA。在17例未治疗的患者中,观察到血清与NT肝脏样本之间丙型肝炎病毒RNA呈正相关且具有统计学意义(Pearson相关系数:rho = 0.544,p = 0.024)。3例接受DAA治疗的患者血清中丙型肝炎病毒RNA“检测不到”,但在所有检测的肝组织中仍“可检测到”。不同的是,仅1例接受DAA治疗且在持续病毒学应答后接受检测的患者,其肝组织中丙型肝炎病毒RNA“检测不到”。

结论

即使血清中丙型肝炎病毒RNA“检测不到”,在肝活检样本和肝外区域中也能成功且高度可靠地定量丙型肝炎病毒RNA。只要有活检样本,雅培实时丙型肝炎病毒检测法就是肝内和肝外区域丙型肝炎病毒定量的良好诊断工具。

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