实时荧光定量RT-PCR法检测手足口病患儿大便标本中肠道病毒71型  

Quantification of enterovirus 71 by real-time fluorescence quantitative RT-PCR in hospitalized children with hand-foot-mouth disease

作　　者：刘丽艳[1] 叶颖子[1] 王建设[1] 俞蕙[1] 朱启镕[1] 

机构地区：[1]复旦大学附属儿科医院传染病研究室,上海200032

出　　处：《临床儿科杂志》2009年第2期142-145,共4页Journal of Clinical Pediatrics

基　　金：上海市卫生系统重点学科建设项目(No.08GWZX0102)

摘　　要：目的了解2008年手足口病流行期间住院手足口病患儿是否存在EV71病毒感染,并探讨实时荧光定量RT-PCR法对手足口病患儿大便标本中EV71病毒载量进行定量检测的可行性。方法采用RT-PCR荧光探针体外扩增法对47例住院的手足口病患儿大便标本中抽提的RNA进行抽样检测。结果22例(46.81%)手足口病患儿大便标本中EV71的病毒载量大于103copies/ml,最高者达1.03×107copies/ml。实时荧光定量RT-PCR法其标准曲线显示,Ct值与病毒拷贝数的对数(log10)之间的相关系数为1.000,相关性较好。结论2008年手足口病流行期间入住儿科医院的手足口病患儿近半数为EV71病毒感染。实时荧光定量RT-PCR法对大便标本中的EV71RNA定量检测较方便快捷,结果直观,为进一步研究病毒载量与临床表现之间的关系打下了基础。Objectives To investigate the hospitalized children with hand-foot-mouth disease caused by enterovirus 71 （EV71） and to develop a real-time fluorescence quantitative reverse transcription-polymerase chain reaction （RT-PCR） for detection of EVT1. Methods EV71 RNA was extracted from the stools of hospitalized children with hand- foot-mouth disease. The extracted RNA was tested by real-time fluorescence quantitative RT-PCR. Results The positive rate of hand-foot-mouth disease caused by EV71 was 46.81% （22/47）. The standard curve of the real-time fluorescence quantitative RT-PCR showed good correlation coefficient （r = 1.000） between the cycle threshold values and logarithm of the viral copy number. Conclusions In this study, 46.81% of the hospitalized children with hand-foot-mouth disease were caused by EV71. Real-time fluorescence quantitative RT-PCR offers a rapid and simple method to detect EV71 from clinical specimens, and will allow to analyze the associations of the clinical symptoms and virus load.

关 键 词：手足口病 肠道病毒71型 实时荧光定量RT—PCR 病毒载量 

分 类 号：R725.1[医药卫生—儿科]