全自动核酸提取工作站对人乳头瘤病毒分型检测的可行性

Study on the feasibility of human papilloma virus subtypes detection by automatic nucleic acid extraction workstation

目的 验证AUTRAX全自动核酸提取工作站对人乳头瘤病毒（HPV）分型检测的可行性。方法 收集2014年8月北京协和医院HPV分型检测样本183份（2 562个型别），应用高危型HPV分型（16、18、35、39、58、31、33、68、56、45、59、51、52）和6、11型核酸测定试剂盒（实时荧光PCR法）进行检测。每份样本分成2份，1份手工提取核酸，配制PCR体系，加样到PCR板；另1份通过AUTRAX全自动核酸提取工作站（以下简称AUTRAX）提取核酸，仪器自动加PCR试剂和提取好的核酸到PCR板，然后均进行实时荧光PCR检测。以手工检测结果为金标准，进行Kappa一致性分析。同时，对AUTRAX检测样本的精密度和防污染进行验证。结果 AUTRAX检测HPV的敏感度为97.12% （101/104），特异度为99.51%（2 446/2 458），准确度为99.42%（2 547 / 2 562）。Kappa一致性结果显示，两种方法高度一致（Kappa＝0.926）；各型别变异系数（CV）均〈5%；无携带污染现象。结论AUTRAX全自动核酸提取工作站可用于临床进行HPV分型检测。

ObjectiveTo verify the feasibility of human papilloma virus（HPV） subtypes detection by AUTRAX automatic nucleic acid extraction workstation.MethodsA total of 183 HPV test samples （2 562 types） were collected during August 2014 in Peking Union Medical College Hospital. Nucleic acid determination kit of high-risk HPV types （16, 18, 35, 39, 58, 31, 33, 68, 56, 45, 59, 51, 52） and 6 , 11 type （real-time PCR） were applied for detection. Each sample was divided into two parts. One part was treated with manual extraction, which entailed manually preparing PCR reaction system and added the sample to the PCR plate. Another was treated with the AUTRAX automatic nucleic acid extraction workstation, which automatically prepared the reaction system and added to the PCR board. These two parts proceeded to the real-time PCR detection. The result of manual extraction was set as the golden standard and the Kappa consistency analysis was conducted. Meanwhile, precision and pollution prevention of the AUTRAX were verified.ResultsThe sensitivity of AUTRAX automatic nucleic acids extraction workstation was 97.12% （101/104）. The specificity was 99.51% （2 446/2 458）. The accuracy was 99.42% （2 547/2 562）. The result of Kappa consistency analysis showed that the two parts were the same （Kappa=0.926）. Coefficient of variation （CV） of each HPV types was less than 5%.No pollution phenomenon was found.ConclusionAUTRAX automatic nucleic acids extraction workstation can be used for the HPV subtypes detection in clinical settings.