Daniel Hubert Darius J, Grant Paul R, Garson Jeremy A, Tedder Richard S, Chandy George M, Abraham Priya
Department of Clinical Virology, Christian Medical College, Vellore-632 004, India.
Diagn Microbiol Infect Dis. 2008 Aug;61(4):415-20. doi: 10.1016/j.diagmicrobio.2008.04.001. Epub 2008 May 16.
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.
即使使用最先进的第三代检测方法，丙型肝炎病毒（HCV）的血清学窗口期约为74天。检测HCV RNA可降低在此期间的传播风险。此外，HCV RNA定量对于合理规划治疗、监测疾病进展以及评估对抗病毒治疗的反应是必要的。我们已经标准化了一种内部HCV实时逆转录聚合酶链反应（RT-PCR）方法，用于筛查以及准确定量和检测血浆样本中的HCV RNA。使用100名慢性感染者和70名乙肝表面抗原、HCV抗体及HIV抗体均为阴性的献血者，将这种内部实时检测方法与一种商业检测方法进行了比较。根据世界卫生组织（WHO）标准评估，这种内部HCV实时RT-PCR的检测下限为50 IU/mL。批间和批内变异系数分别为1.3%至6.4%和0.0%至2.3%。用这种内部HCV实时检测方法估计的病毒载量与商业的artus HCV RG RT-PCR检测方法相关（r = 0.59，P < 0.0001）。该检测方法可用于筛查和监测接受治疗的个体，在检测中未显示出基因型依赖性差异。
