摘要
为建立一种可以快速、准确检测O型口蹄疫病毒的TaqMan荧光定量PCR方法,根据O型口蹄疫病毒VP1基因保守区序列设计特异性引物和TaqMan探针,使用MagMax Express核酸自动提取系统提取O型口蹄疫病毒FMD/O/YM/YN/2000株病毒核酸,建立了检测O型口蹄疫病毒的实时荧光定量PCR方法。该方法标准曲线相关系数达0.999 3,最低检测量达到1.79个拷贝,比传统荧光定量PCR方法灵敏10倍以上,特异性强。该方法可以在2.5h内检测96份临床样品,相对于传统荧光定量PCR方法,效率提高2倍以上。采用MagMax Express核酸自动提取系统配合7500Fast real-time PCR system建立的方法可以实现快速、准确检测O型口蹄疫病毒。
This research established a real-time PCR method based on 1 aqman probe tnar coulct ueL^ct ~ virus type O rapidly and accurately. We used ABI Magmax Express massive flux automatic nucleic acid ex- traction system to extract FMD virus type O strain FMD/O/YM/YN/2000, and designed specific primers and Taqman probe according to FMD virus type O VP1 gene conservative zone, then established the meth- od of massive flux real-time PCR for detection of FMD type O. The result of standard curve of this method is very accurate, and the relative coefficient is 0. 999 3, and the lowest detection limit is 1.79 copies, which is over 10 times more sensitive than traditional real-time PCR method, and this method is specific. We de- tected 96 clinical samples within 2.5 hours, which only cost up to half time of traditional methods. This research proved that using ABI MagMax Express massive flux automatic nucleic acid extraction system with ABI 9700 fast real time PCR system could detect FMD virus type O in massive flux, rapidly and accu- rately.
出处
《动物医学进展》
CSCD
北大核心
2014年第1期50-53,共4页
Progress In Veterinary Medicine
基金
农业部公益性行业(农业)科研专项(201103008)
