逆转录-荧光实时定量PCR法快速测定肠道病毒71型病毒滴度  

Quantitative reverse transcriptase PCR for quick titer determination of enterovirus 71

作　　者：李剑波[1] 吴洁[1] 高丽美[1] 周康凤[1] 毛卓玥 高孟[1] 庄昉成[1] 

机构地区：[1]浙江省医学科学院病毒病研究所,杭州310013 [2]杭州市第十四中学,310006

出　　处：《国际流行病学传染病学杂志》2017年第5期312-316,共5页International Journal of Epidemiology and Infectious Disease

基　　金：浙江省医学重点学科;浙江省科技计划（2017C37120）;浙江省中医药科技计划（2017ZA038）

摘　　要：目的 建立一种肠道病毒71型（EV71）病毒滴度快速测定法.方法 以TCID50法测定的EV71病毒样品作为滴度检测参考品,建立EV71病毒滴度的逆转录-荧光实时定量PCR测定方法.通过滴度参考品和待测病毒样品的Ct值计算待测样品中的病毒滴度,并对该检测法进行特异性、重复性验证.结果 所得检测方法可以用于EV71病毒滴度的快速检测.本测定法具有较好的线性,线性范围为lgTCID50 2.61-8.61,相关系数R2为0.9881;特异性和重复性较好,与TCID50法检测之间具有较好的一致性,变异系数（CV）＜5％.结论 所建立的逆转录-荧光实时定量PCR法线性范围广,具有较好的重复性和特异性,可用于EV71样品病毒滴度的快速测定.Objective To develop a quick-convenient method for the titer determination of enterovirus 71 （EV71）.Methods The traditional method TCID50 was used to determine the sample titer of EV71 as a reference.The reference and other samples were re-tested by quantitative RT-PCR.The EV71 titer was calculated by Ct values of the reference and EV71 samples.The specificity and repeatability of this method was verified.Results The detecting approach could be used for quick-convenient titer detection of EV71.RT-PCR showed a good linearity with the testing virus titer lgTCID50 range of 2.61-8.61 and correlation coefficient （R2） of 0.9881.The method had good specificity and repeatability,and was well consistent with TCID50 method with the coefficient of variation （CV）〈5%.Conclusions The developed quantitative RT-PCR method shows a wide linear range,high specificity and superior repeatability.It can be a quick-convenient method for titer determination of EV71.

关 键 词：逆转录聚合酶链反应 肠道病毒71型 病毒滴度 

分 类 号：R446.5[医药卫生—诊断学]