基因分型质控品在评价PCR-杂交法人乳头瘤病毒基因分型检测试剂中的应用

Evaluation of PCR-hybridization human papillomavirus genotyping kit by genotyping quality control materials

目的:使用人乳头瘤病毒L1基因分型质控品对PCR-杂交法人乳头瘤病毒基因分型检测试剂盒进行评价。方法:采用建立的HPV基因分型质控品,以质控品盘包含的30种型别的质粒DNA为样本,按照各个试剂盒要求进行扩增、杂交、检测。结果:检测HPV基因分型质控品盘30种不同型别HPV质粒,试剂A检测30种型别检测结果全部符合;试剂B检测有28种型别检测结果符合,2种型别不符合,HPV26和68型漏检;试剂C检测有28种型别检测结果符合,2种型别不符合,HPV44和68型漏检;试剂D检测有27种型别检测结果符合,3种型别不符合,HPV26,44和73型漏检;4家公司的试剂盒基本能正确区分质控品中HPV基因型,但有部分的HPV基因型有漏检现象。结论:包含30种不同型别的HPV基因分型质控品能可靠评价4家公司的HPV分型试剂盒(PCR-杂交法);试剂盒的研发应在通过质控盘的考核情况下加大临床样本的验证,同时也应大力开展HPV分子流行病学的研究,为试剂盒的研发储备基础资料。

Objective：To evaluate the PCR-hybridization human papillomavirus（HPV） genotyping kit by the HPV L1-gene genotyping control materials.Methods：The HPV L1-gene genotyping control materials including 30 different HPV types plasmids were amplified,hybridized and detected as samples,according to the kit instructions.Results：The HPV L1-gene genotyping control materials including 30 different HPV types plasmids as template,for kit A ,30 HPV types plasmids were accurately genotyped;for kit B ,28 HPV types plasmids were accurately genotyped,and two types were wrong which HPV type 26 and 68 were missed;for kit C ,28 HPV types plasmids were accurately genotyped,and two types were wrong which HPV type 44 and 68 were missed;for kit D ,27 HPV types plasmids were accurately genotyped,and three types were wrong which HPV type 26,44 and 73 were missed.For the four kits,the HPV genotypes detection was close to the right,but some HPV types had been missed.Conclusion：The four PCR-hybridization HPV genotyping kits were effectively evaluated by the HPV L1-gene genotyping control materials.The kit development should be verified by large scale clinical samples after HPV genotyping control materials evaluation.Furthermore,the molecular epidemiology of HPV should be focused on and research data was used for the kit development.