摘要目的 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).结论

abstractsObjective 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.