实时荧光PCR技术在丙型肝炎病毒基因型检测中的应用

Application of real-time fluorescent PCR in the detection of HCV genotypes.

目的探讨实时荧光PCR技术在丙型肝炎病毒(HCV)基因型检测中的应用价值。方法采用一步法逆转录聚合酶链反应(RT-PCR)结合TaqMan技术,对HCV基因型进行实时荧光PCR检测;巢式RT-PCR扩增HCV的NS5B基因区域,测序后进行HCV基因亚型分析并与实时荧光PCR分型结果进行比较。结果实时荧光PCR共分型119例,1型71例(59.7%),非1型48例(40.3%)其中110例成功进行测序分型;测序分型的110例中,1型73例(均为1b型),占66.4%,非1型37例(其中2a型7例、3a型6例、3b型3例、4a型1例、4k型1例、5a型1例、6a型17例、6n型1例),占33.6%。与110例测序分型的结果比较,实时荧光PCR分型总的符合率为71.8%(79/110),其中1型的符合率为80.9%(55/68),非1型的符合率为57.1%(24/42)。结论实时荧光PCR进行HCV基因型检测操作简便、快速,但分型结果特异性不高。

Objective To study the application of real-time fluorescent PCR in the detection of Hepatitis C Virus（HCV） genotypes.Methods Real-time fluorescent PCR with one step reverse transcription（RT） combining with TaqMan was performed to detect the HCV genotypes.And sequencing of NS5B gene was applied for comparison.Results Real-time fluorescent PCR genotyping was successfully performed in 110 of 119 cases of HCV,with 71 and 48 cases were genotype-1 and non-genotype-1,respectively.Genotype-1 and non-genotype-1 were revealed in 73 cases（66.4%,all were subtype 1b） and 37 cases（33.6%,7 cases were 2a,6 cases were 3a,3 cases were 3b,1 case was 4a,1 case was 4k,1 case was 5a,17 cases were 6a and 1 case was 6n）,respectively.The total coincidence rates of genotype-1 and non-genotype-1 were 71.8%（79/110 and 80.9%（55/68）,respectively.Conclusion Real-time fluorescent PCR is rapid and simple test for HCV genotyping,though without high specificity.