HCV基因分型试剂盒评价血清盘的构建与应用  

Establishment and application of evaluation serum panel for hepatitis C virus genotyping kit

作　　者：陈兵[1] 马仲慧 常浩[1] 裴丽健[1] 蒋岩[1] 邱茂锋[1] 肖瑶[1] 邢文革[1] 

机构地区：[1]中国疾病预防控制中心性病艾滋病预防控制中心参比室,北京102206

出　　处：《中国生物制品学杂志》2017年第5期505-508,共4页Chinese Journal of Biologicals

摘　　要：目的构建HCV基因分型试剂盒评价血清盘,对国内HCV基因分型试剂盒进行评价与应用,以评估我国HCV基因分型试剂盒的质量。方法收集全国不同省份、不同人群疑似HCV的样本,首先对样本进行筛查试验和病毒载量检测,选取样本的病毒载量大于1 000 IU/ml,再用测序法确定样本的HCV基因型,对确定基因型的样本进行分析,每个基因亚型选择2支,再加上2支病毒载量为阴性的样本,构建成1套HCV基因型试剂盒评价盘,分装并评价其稳定性,并利用评价盘对HCV基因分型试剂盒进行质量评估。结果 HCV NS5B区、CORE区测序基因型结果与血清盘参考预期结果的准确性均为100%(7/7);828份样本中共检测出7种基因亚型,分别为1a、1b、2a、3a、3b、6a、6n,未检测到其他罕见型别(4型、5型),综合考虑各方面因素,每个基因亚型选择2支样本再加上2支HCV阴性样本共16支样本,组成HCV基因型试剂盒评价盘;经反复冻融3次后检测基因型,其稳定性无影响;利用构建的HCV基因型评价盘对分型试剂盒进行评价,其基因型检出准确性为9/14,其中试剂盒检测范围之内的5种型别有1个样本未检测(3b),而对于检测范围之外的基因型(1a、6n),将基因型1a均错判为1b(2/2)。结论本实验室构建的HCV基因分型评价盘充分考虑了我国HCV基因型的流行情况及国内HCV基因分型试剂盒的检测范围,能够对国内HCV基因分型试剂盒的质量进行有效评价。Objective To establish evaluation panel for hepatitis C virus （HCV） genotyping kit and evaluate the quality of domestic HCV genotyping kit.Methods Suspected HCV samples from various populations across different provinces in China were collected for screening test and determination of viral load.The samples with viral load of more than 1 000 IU/ml were determined for HCV genotype by sequencing,anid those of defined genotypes were analyzed.Two samples of each subtype plus samples negative for viral load were selected to establish a set of HCV genotyping kit evaluation panel,which was filled,evaluated for stability,and used for quality evaluation of HCV genotyping kit.Results All the accuracies of genotype results of HCV NSSB region,CORE region and reference expected results of serum panel were 100% （7/7）.Seven gene subtypes were detected from 828 samples,including 1a,1b,2a,3a,3b,6a and 6n,while no other rare types （4 or 5） were detected.Considering all the factors,a HCV genotype kit evaluation panel was established by using two samples of each gene subtype plus two HCV negative samples,16 in total.Three cycles of repeated freezing and thawing showed no effect on the stability of the panel.The accuracy of the panel for detection of genotype was 9 / 14 in evaluation of HCV genotyping kit.Genotype 3b as one of the five genotypes within the detection scope of the kits was not detected.However,for genotypes beyond the detection scope,such as 1a and 6n,both the samples of genotype 1a were judged as of 1b.Conclusion The homemade HCV genotyping evaluation panel in our laboratory gives full consideration to the prevalence of HCV genotypes in China and detection range of domestic HCV genotyping kit,which contributes the effective evaluation of domestic HCV genotyping kit.

关 键 词：丙型肝炎病毒 基因分型试剂盒 血清盘 

分 类 号：R373.21[医药卫生—病原生物学] R392-33[医药卫生—基础医学]