Real-Time RT-PCR检测手足口病肠道病毒方法的建立及应用

Establishment and Application of Real-time RT-PCR Method for Detection of Enteroviruses of Hand,Foot and Mouth Disease

目的建立手足口病肠道病毒的Real-Time RT-PCR法,为手足口病病毒快速、准确检测提供一种新方法。方法采用卫生部《手足口病预防控制指南》(2009年版,以下简称《指南》)推荐的RT-PCR法对湖南省株洲市2009年3-8月间发生的82例临床诊断手足口病患儿的82份标本进行人肠道病毒、柯萨奇病毒A16型和肠道病毒71型特异性核酸的检测,同时采用实时荧光定量RT-PCR(Real-Time Quantitative RT-PCR,简称Real-TimeRT-PCR)进行复检,比较二者的实验结果 ,分析Real-TimeRT-PCR的准确度和灵敏度。结果 Real-TimeRT-PCR检测82份标本肠道病毒、柯萨奇病毒A16型、肠道病毒71型,与RT-PCR比较,二者差异有统计学意义(λ=4.09,P<0.05),Real-TimeRT-PCR法检出率明显高于RT-PCR法。结论本研究建立了一种灵敏、特异、简单易行的Real-TimeRT-PCR检测手足口病肠道病毒的方法 ,为手足口病病毒的快速诊断提供了新的方法 。

Objective To establish the Real-time RT-PCR method for hand-foot-and-mouth disease （HFMD） and to provide a quick and accurate testing way for HMFD.Methods According to recommendation from the Guidelines of Hand-foot-and-mouth Disease Prevention （2009）,Real-time RT-PCR method was used to detect human enterovirus,coxsackievirus A16,and enterovirus 71 in 82 HFMD cases clinically diagnosed from March to August,2009 in Zhuzhou.In addition,Real-time Quantitative RT-PCR was used to retest the cases.The results of the two tests were compared,and the accuracy and sensitivity of Real-time RT-PCT were analyzed.Results There were statistically significant differences in detection of human enterovirus,coxsackievirus A16,and enterovirus 71 in 82 HFMD cases between Real-time RT-PCT and RT-PCT （λ=4.09,P〈0.05）.Real-time RT-PCR was more sensitive than RT-PCR.Conclusions This study has established a sensitive,easy and quick Real-time RT-PCR method for the detection of HFMD which provides a new way for quick test of HFMD.