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.
出处
《黑龙江畜牧兽医》
CAS
北大核心
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
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
recombinase polymerase amplification
reverse transcription real - time recombinase polymerase amplification
Foot - and - mouth disease virus （FMDV）
rapid detection
