Department of Surgical Gastroenterology, Bhopal Memorial Hospital & Research Center, Karond, Raisen Bypass Road, Bhopal, 462 038, Madhya Pradesh, India.
J Gastrointest Surg. 2012 Jan;16(1):142-6; discussion 146-7. doi: 10.1007/s11605-011-1750-0. Epub 2011 Nov 3.
Hepatitis C virus (HCV) infects nearly 3% of the population worldwide and is a major cause of acute and chronic infections leading to fibrosis, cirrhosis, and hepatocellular carcinoma. Current laboratory diagnosis of HCV is based on specific antibody detection (anti-hepatitis C virus (anti-HCV)) in serum. As HCV replicates in the liver cells, detection and localization of HCV RNA in liver tissue are vital for diagnosis.
Ten biopsy samples diagnosed for cryptogenic liver cirrhosis, negative for the presence of anti-HCV and serum HCV RNA, were studied for analyzing presence of viral nucleic acid in liver tissues. Qualitative screening for HCV was done through ELISA while the nucleic acid analysis was performed through COBAS Amplicor. Detection of HCV RNA in liver tissue biopsies was performed following standard protocol of HCV detection kit (Shenzhen PG Biotech) with modifications using Light Cycler 2.0 (minimum detection limit 10 copies/ml).
Quantitative detection in liver biopsies following the modified method showed the presence of HCV RNA in three samples out of the ten studied.
The results indicate that using Light Cycler 2.0, following the modified technique described, constitutes a reliable method of quantitative detection and localization of HCV in tissue in "serosilent" HCV infection.
丙型肝炎病毒 (HCV) 在全球范围内感染了近 3%的人口，是导致纤维化、肝硬化和肝细胞癌的急性和慢性感染的主要原因。目前 HCV 的实验室诊断基于血清中特异性抗体的检测（抗丙型肝炎病毒 (抗-HCV)）。由于 HCV 在肝细胞内复制，因此检测和定位肝组织中的 HCV RNA 对于诊断至关重要。
对诊断为不明原因肝硬化、抗-HCV 和血清 HCV RNA 均为阴性的 10 个活检样本进行研究，以分析肝组织中是否存在病毒核酸。通过 ELISA 进行 HCV 的定性筛选，通过 COBAS Amplicor 进行核酸分析。按照 HCV 检测试剂盒（深圳 PG 生物技术有限公司）的标准方案进行肝组织活检中 HCV RNA 的检测，同时采用 LightCycler 2.0 进行修改（最小检测限为 10 拷贝/ml）。
经改良方法对肝活检进行定量检测，在 10 个研究样本中有 3 个样本中检测到 HCV RNA。
这些结果表明，使用 LightCycler 2.0，采用描述的改良技术，构成了一种在“血清沉默”HCV 感染中定量检测和定位 HCV 的可靠方法。
