水疱性口炎病毒荧光RT-PCR鉴别检测方法的建立与应用  

Establishment and Application of a Taq Man Real-time RT-PCR Assay for the Detection of Vesicular Stomatitis Virus

作　　者：臧京帅[1] 高志强[2] 王丽萍[1] 郭金玉[1] 傅毅 牛建蕊 刘文晓 张乐萃[1] 张鹤晓[2] 

机构地区：[1]青岛农业大学动物科技学院,山东青岛266109 [2]北京出入境检验检疫局,北京朝阳100026 [3]北京森康生物技术开发有限公司,北京怀柔101400

出　　处：《中国兽医杂志》2015年第5期73-76,共4页Chinese Journal of Veterinary Medicine

基　　金：质检公益性行业科研专项"马重大外来虫媒病抗体;核酸快速检测技术研究"(201010033)

摘　　要：本研究建立荧光定量RT-PCR鉴别检测水疱性口炎病毒的方法。针对Gen Bank登录的水疱性口炎两血清型-新泽西型(VSV-NJ)和印第安纳型(VSV-IND)的L基因保守区,设计1对引物和2条探针。通过对荧光定量RT-PCR反应条件的优化,建立了Taq Man荧光定量RT-PCR快速检测VSV的方法。与常规RT-PCR试验比较表明,所建立的荧光RT-PCR检测技术快速、敏感,检测时限3 h以内,具有很好的特异性和重复性,可以实现水疱性口炎病毒的快速检测分型。通过对342份临床样品进行检测,结果表明,所建立的检测方法适用于样品中水疱性口炎病毒的直接检测。The study aimed to develop qRT-PCR methods for the identification and detection of Vesicular Stomatitis Virus. The probes and primers were designed and synthesized based on the L gene conserved region of Two Types of Vesicular Stomatitis Virus （ VSV-NJ and VSV-IND ） , which were available in GenBank. The reaction parameters were optimized, and a sensitive Taq- Man-based Real-time PCR assay for the rapid detection of VSV was developed. Compared with normal RT-PCR, the developed qRT-PCR was fast and sensitive, and the detection time was shortened to about 3 hours. Furthermore, the detection method showed great specificity and reproducibility. By detecting 342 clinical samples, the results suggested that the method could be used to de- tect VSV from samples directly.

关 键 词：水疱性口炎病毒 荧光RT-PCR TCID50 鉴别检测 

分 类 号：S854.43[农业科学—临床兽医学]