PCR-荧光探针法在中国HCV常见基因型检测中的意义

Application of PCR-fluorescent probe in identifying genotypes of HCV

目的 探讨聚合酶链反应（Polymerase chain reaction,PCR）-荧光探针法在检测丙型肝炎病毒（HCV）基因型中的应用价值.方法 随机选择2016年3月至6月来自全国各地的166例慢性HCV感染者血清样本,采用逆转录巢式PCR扩增HCV的Core-E1（CE1）基因区域和测序、系统发育树分析确定基因亚型,并采用PCR-荧光探针法对HCV基因型进行检测.采用Kappa检验对两种方法测得的HCV分型结果进行比较.结果 巢式PCR测序法共分型166例,其中1 b型66例（39.8%）,2a型34例（20.5%）,3a型16例（9.6%）,3b型为27例（16.2%）,6a型23例（13.9%）;PCR-荧光探针法共分型166例,其中1 b型68例（41.0%）,2a型34例（20.5%）,3a型16例（9.6%）,3b型25例（15.0%）,6a型23例（13.9%）.PCR-荧光探针法分型中的2例1 b型,测序法分型为3b型,两种方法检测一致率为98.7%（164/166）,差异无统计学意义（χ2=0.0492,P〉0.05）.

Objective To evaluate the application value of polymerase chain reaction （PCR）-fluorescence probe method in identifying genotypes of hepatitis C virus （HCV）.Methods One hundred and sixty six serum samples from patients with chronic HCV infection were collected nationwide from March to June 2016.HCV Core-E1 gene region was amplified and sequenced by nested reverse transcription-PCR （RT nested-PCR）and genetic subtypes were analyzed by phylogenetic tree,meanwhile HCV genotypes were also determined by PCR-fluorescent probe method.Kappa test was used to compare the consistency of two methods.Results Among 166 samples detected by RT nested-PCR,the genotype of 66 samples （39.8%） was 1 b,34 （20.5%）was 2a,16 （9.6%）was 3a,27 was 3b （16.2%）,23 （13.9%）was 6a.Two samples with 3b genotype detected by RT nested-PCR were identified as 1 b by PCR-fluorescent probe.The consistency rate of two methods was 98.7% （164 /166）,there was no significant difference between two methods （χ2 =0.0492,P 〉0.05）.Conclusion PCR-fluorescence probe method can accurately identify HCV genotypes and can be used in clinic.