鸭坦布苏病毒E基因环介导等温扩增检测方法的建立与应用  

Development and application of a loop-mediated isothermal amplification assay for detection of Tembusu virus E gene

作　　者：欧全宾[1] 刁有祥[1] 唐熠[1] 高绪慧[1] 于春梅[1] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018

出　　处：《中国兽医学报》2013年第9期1329-1332,共4页Chinese Journal of Veterinary Science

基　　金：现代农业产业技术体系建设专项基金资助项目(CARS-43-34)

摘　　要：根据GenBank发布的坦布苏病毒Bagaza毒株E基因的保守序列设计1套引物,建立了环介导等温扩增快速检测鸭坦布苏病毒的方法。该检测方法的敏感性可达10拷贝/μL;对鸭坦布苏病毒山东分离株的检测呈阳性,而对鸭瘟病毒、禽流感病毒(H9亚型)、鸭新城疫病毒、传染性法氏囊病病毒、减蛋综合征病毒扩增结果均为阴性;应用该方法对山东省各地采集的82份疑似病料进行检测,可检出72份阳性。表明本试验所建立的环介导等温扩增检测方法灵敏性高、特异性强、操作简便快速,可用于鸭坦布苏病毒病的临床诊断和流行病学调查。A loop-mediated isothermal amplification（LAMP）assay was developed for the rapid detection of Tembusu virus（TMUV）using a set of specific primers targeting on the E gene conserved region of TMUV Bagaza strain published in GenBank.The assay has the lowest detection limit being 10 copies/μL,and one strain of TMUV of Shandong isolates were positive,and there was no cross-reactivity with duck plagues virus（DPV）,avian influenza virus（AIV）subtype H9,Newcastle disease virus（NDV）,infectious bursal disease virus（IBDV）and egg drop syndrome virus（EDS-76）.Application of the method,82 suspicious samples collecting from Shandong province was tested and 72 samples was positive.These results suggested that the LAMP assay with high sensitivity,strong specificity,simple operation could be used as a method for the diagnosis and detection of clinical cases,and molecular epidemiological investigation of duck TMUV.

关 键 词：坦布苏病毒 E基因 环介导等温扩增 荧光染料 

分 类 号：S852.65[农业科学—基础兽医学]