FMDV、BTV、PPRV多重RT-PCR检测方法的建立
Establishment of Multiplex RT-PCR Method for Detecting FMDV, BTV and PPRV

为建立能够同时检测并鉴别口蹄疫病毒（Foot-and-mouth disease virus，FMDV）、蓝舌病病毒（Bluetongue virus，BTV）和小反刍兽疫病毒（Peste des petits ruminants virus，PPRV）感染的多重RT-PCR（反转录-聚合酶链式反应，Reverse transcription-polymerase chain reaction）方法，根据3种病毒的基因组全序列和保守区序列设计 3 对特异性引物，优化反应体系和扩增条件，建立能够同时检测 3 种病毒的多重RT-PCR方法。结果表明，建立的多重RT-PCR检测方法敏感性强，对FMDV、BTV和PPRV的核酸最低检测量分别为15.42、6.29、16.50 ng/μL；特异性试验结果表明所建立方法的特异性良好。建立的多重RT-PCR方法具有敏感性高、特异性强、重复性好、快速简便等特点，可以用于临床上 3 种病毒感染的诊断和鉴别。
In order to develop multiplex RT-PCR method for the clinical detection of FMDV, BTV and PPRV infection of goats and sheep, three pairs of primers were designed and synthesized according to the highly conserved regions of FMDV, BTV and PPRV genome sequence in GenBank. The multiplex RT-PCR reaction condition was optimized and the multiplex RT-PCR method for detecting FMDV, BTV and PPRV infection was established. The results showed that the newly-established method had high sensitivity. The lowest levels of the three viruses were 15.42 ng/μL,6.29 ng/μL and 16.50 ng/μL, respectively. The specificity test showed that the newly-established method has high specificity. The result indicated multiplex RT-PCR method had specificity, sensitivity, repetitive characteristics and it has applied value in detection of FMDV, BTV and PPRV infection．