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| 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 | |
| 引用本文： | 徐丽秋,苏永生,赵玉军,刘海防,高玉峰,王长文,贾赟.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[J].畜牧与兽医,2009,41(9). |
| 作者姓名： | 徐丽秋 苏永生 赵玉军 刘海防 高玉峰 王长文 贾赟 |
| 作者单位： | 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 |
| 基金项目： | 辽宁出入境检验检疫局科技项目资助 |
| 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。 |
| 关 键 词： | 猪传染性胃肠炎病毒(TGEV) 实时定量RT-PCR TaqMan荧光探针 |
Establishment and preliminary application of a TaqMan real-time RT-PCR method for detection of porcine transmissible gastroenteritis virus |
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| XU Li-qiu,SU Yong-sheng,ZHAO Yu-jun,LIU Hai-fang,GAO Yu-feng,WANG Chang-wen,JIA Yun.Establishment and preliminary application of a TaqMan real-time RT-PCR method for detection of porcine transmissible gastroenteritis virus[J].Animal Husbandry & Veterinary Medicine,2009,41(9). | |
| Authors: | XU Li-qiu SU Yong-sheng ZHAO Yu-jun LIU Hai-fang GAO Yu-feng WANG Chang-wen JIA Yun |
| Abstract: | 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 RT-PCR 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. |
| Keywords: | transmissible gastroenteritis virus(TGEV) real-time quantitative RT-PCR TaqMan fluorescence probe |
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