口蹄疫病毒real-time RT-RPA检测方法的建立与应用  

Establishment and application of real-time RT-RPA for rapid detection foot-and-mouth disease virus

作　　者：杨洋[1] 秦晓东[1] 宋一鸣[1] 施冲旭 李彦敏[1] 张志东[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,兰州730046

出　　处：《黑龙江畜牧兽医》2017年第5期172-176,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：中国农业科学院创新团队项目

摘　　要：为了建立口蹄疫病毒(FMDV)的快速检测方法,试验以FMDV 3D基因作为靶基因设计引物和探针,建立了一种新的实时荧光反转录重组酶聚合酶扩增(real-time RT-RPA)检测方法,该方法在40℃恒温条件下20 min内完成检测,并评估了该方法的特异性、敏感性和重复性。结果表明:该检测方法能特异性检测FMDV,敏感性为500拷贝/反应,而且具有很好的重复性。利用该方法对采集的临床样品进行检测,检测结果与RT-q PCR检测结果具有100%的符合率。说明real-time RT-RPA方法具有简单、快速、特异性强的优点,是一种潜在的FMDV快速检测方法。To establish a method for rapid detection of Foot - and - Mouth Disease Virus （ FMDV）, a novel fluorescent probe - based reverse transcription recombinase polymerase amplification （real -time RT - RPA） was developed by using primer and probe specificaUy targeting the 3D gene of FMDV. The detection of the real - time RT - RPA could be finished in 20 rain at 40 ~C. Furthermore, the real - time RT - RPA was subjected to evaluation of sensitivity, specificity and reproducibility. The results showed that the real - time RT - RPA exhibited high speci- ficity for detection of FMDV ; sensitivity of the real - time RT - RPA was as low as 500 copies of FMDV RNA/reaction; the real - time RT - RPA had a good reproducibility. Results of the clinical detection showed that the developed real - time RT - RPA had 100% consistency when compared with RT - qPCR assays. These data demonstrated that the developed real - time RT - RPA had a potential use for rapid detection of FMDV.

关 键 词：重组酶聚合酶扩增 实时荧光反转录重组酶聚合酶扩增 口蹄疫病毒(FMDV) 快速检测 

分 类 号：S852.65[农业科学—基础兽医学] Q814.9[农业科学—兽医学]