论文部分内容阅读
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 construct a HCV genotyping kit for the evaluation of the serum disk and to evaluate and apply the domestic HCV genotyping kit to evaluate the quality of the HCV genotyping kit in China. Methods The samples of suspected HCV in different provinces and different populations in China were collected. The samples were firstly screened and the viral load was tested. The viral load of samples was more than 1 000 IU / ml. The HCV genotype was determined by sequencing. Two genotypes of each genotype were selected for analysis of genotyped samples, plus two samples of negative viral load to construct a suite of HCV genotyping kits for packaging and evaluation of their stability , And evaluated the quality of the HCV genotyping kit using the evaluation panel. Results The accuracy of sequencing results of genotypes of HCV NS5B and CORE was 100% (7/7) with reference to the expected result of serological disk analysis. Seven genotypes were detected in 828 samples, which were 1a, 1b, 2a, 3a, 3b, 6a, 6n, and other rare types (type 4 and type 5) were not detected. Considering all the factors, two samples were selected for each gene subtypes plus two samples of HCV negative , Which made HCV genotyping kit evaluation plate. After repeated freezing and thawing three times, the genotypes were tested and the stability was not affected. The genotyping kit was evaluated by the constructed HCV genotype evaluation plate, and the accuracy of genotype detection 9/14, of which 1 sample out of 5 types within the detection range of the kit was not detected (3b), while genotypes 1a and 6n were excluded from the detection range 1b (2/2). Conclusion The HCV genotyping panel constructed in our laboratory fully considers the prevalence of HCV genotypes in China and the detection range of HCV genotyping kits in China and can effectively evaluate the quality of HCV genotyping kits in China.