检测猪传染性胃肠炎病毒Taq Man荧光定量RT-PCR方法的建立与初步应用  

Establishment and preliminary application of a TaqMan real-time RT-PCR method for detection of porcine transmissible gastroenteritis virus

作　　者：徐丽秋[1] 苏永生[2] 赵玉军[1] 刘海防[3] 高玉峰[4] 王长文[2] 贾赟[2] 

机构地区：[1]沈阳农业大学动科院,辽宁沈阳110161 [2]辽宁出入境检验检疫局,辽宁大连116001 [3]山东农业大学动科院,山东泰安271018 [4]大连工业大学生物与食品工程学院,辽宁大连116034

出　　处：《畜牧与兽医》2009年第9期14-17,共4页Animal Husbandry & Veterinary Medicine

基　　金：辽宁出入境检验检疫局科技项目资助(LK12-2007)

摘　　要：据GenBank登录的猪传染性胃肠炎病毒(TGEV)N基因的保守区序列设计合成了引物和探针,对荧光定量RT-PCR的反应条件进行了优化,建立了一种特异、灵敏、快速的TaqMan荧光定量RT-PCR方法来检测TGEV。与国外进口的An imal Genetics公司生产TGEV抗原快速检测试剂盒进行比较,符合率达到100%;与常规RT-PCR检测方法相比,灵敏度高100倍。用该方法对辽宁、山东、福建等地的送检疑似病料进行了检测,结果阳性率为24%。试验证明,所建立方法适用于猪传染性胃肠炎病毒的分子诊断、流行病学调查等相关研究。The probes and primers were designed and synthesized according to the conserved gene N of transmissible gastroenteritis virus （TGEV） available in GenBank, and then reaction parameters were optimized to develop a specific and sensitive TaqMan-based real-time RTPCR assay for the rapid detection of TGEV. Compared with the imported TGEV rapid antigen detection kit from Animal Genetics, the coincidence rate was 100%. The TGEV samples from the swine farms in Liaoning, Shandong and Fujian Province were detected by the established quantitative RT-PCR. The results indicated that 24% was positive. It suggested that the method could be used in clinical diagnosis and epidemiological investigation.

关 键 词：猪传染性胃肠炎病毒(TGEV) 实时定量RT—PCR TaqMan荧光探针 

分 类 号：S852.65[农业科学—基础兽医学]