Quantitation of hepatitis C virus using an in-house real-time reverse transcriptase polymerase chain reaction in plasma samples
Quantitation of hepatitis C virus using an in-house real-time reverse transcriptase polymerase chain reaction in plasma samples
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DOI：
10.1016/j.diagmicrobio.2008.04.001
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发表时间：
2008-08-01
影响因子：
2.9
通讯作者：
Abraham, Priya
中科院分区：
文献类型：
--
作者：
Daniel, Hubert Darius J.;Chandy, George M.;Abraham, Priya
即使采用最先进的第三代检测方法，丙型肝炎病毒(丙型肝炎病毒)的血清学窗口期也约为74天。检测丙型肝炎病毒核糖核酸将降低在此期间传播的风险。此外，定量检测丙型肝炎病毒核糖核酸对于正确的治疗计划、监测疾病进展和评估抗病毒治疗的反应是必要的。我们已经标准化了一种内部的丙型肝炎病毒实时逆转录聚合酶链式反应(RT-PCR)，用于筛选和准确定量和检测血浆样本中的丙型肝炎病毒RNA。对100名慢性感染者和70名乙肝表面抗原、丙型肝炎病毒抗体和艾滋病毒抗体阴性的献血者进行了室内实时检测和商业检测。根据世界卫生组织(WHO)的标准，这种内部丙型肝炎病毒实时RT-PCR的检测下限为50IU/毫升。批间变异系数为1.3%~6.4%，批内变异系数为0.0%~2.3%。用这种内部丙型肝炎病毒实时检测方法估计的病毒载量与商业的ARTUS丙型肝炎病毒RG RT-PCR法相关(r=0.59P<；0.0001)。该检测方法可用于筛查和监测接受治疗的个体，在检测中没有显示出依赖于基因型的差异。(C)2008 Elsevier Inc.保留所有权利。
Even with the most advanced 3rd-generation assays, the serologic window period of hepatitis C virus (HCV) is approximately 74 days. HCV RNA detection would reduce the risk of transmission during this period. Furthermore, quantitation of HCV RNA is necessary for proper planning of treatment, monitoring disease progression, and assessing response to antiviral therapy. We have standardized an in-house HCV real-time reverse transcriptase polymerase chain reaction (RT-PCR) for screening and accurate quantitation and detection of HCV RNA in plasma samples. The in-house real-time assay was compared with a commercial assay using 100 chronically infected individuals and 70 blood donors who are negative for hepatitis B surface antigen, HCV antibody, and HIV antibody. The lower limit of detection of this in-house HCV real-time RT-PCR as assessed against the World Health Organization (WHO) standard was 50 IU/mL. Interassay and intraassay coefficient of variation ranged from 1.3% to 6.4% and 0.0% to 2.3% respectively. Virus loads as estimated with this in-house HCV real-time assay correlated with the commercial artus HCV RG RT-PCR assay (r = 0.59, P < 0.0001). This assay could be used in screening and monitoring individuals on therapy, showing no genotype-dependent differences in detection. (c) 2008 Elsevier Inc. All rights reserved.