传染性支气管炎病毒RT-LAMP检测方法的建立  

Development of an RT-LAMP method for the detection of infectious bronchitis virus

作　　者：范瑾[1] 谭磊[2,3] 仇旭升[2,3] 孟春春[2,3] 包世俊[2,3] 胡美容[2,3] 何随彬[2,3] 郭小琴[2,3] 于圣青[2,3] 陈书明[1] 丁铲[2,3] 

机构地区：[1]山西农业大学动物科技学院,山西太谷030801 [2]中国农业科学院上海兽医研究所,上海200241 [3]国家家禽工程技术研究中心,上海200331

出　　处：《中国兽医科学》2012年第11期1163-1167,共5页Chinese Veterinary Science

基　　金：中央级公益性科研院所基本科研业务费专项资金项目(2012JB11);山西省科技攻关项目(052011);山西高校科技研究开发项目(20051222)

摘　　要：为建立一种快速、灵敏、特异的检测传染性支气管炎病毒(IBV)的逆转录-环介导等温扩增方法(RT-LAMP),针对IBV毒株保守性N蛋白设计4对LAMP特异性引物,以RNA为模板,在Bst DNA聚合酶作用下恒温反应,对不同型IBV、新城疫病毒、禽流感病毒和鸡滑液支原体进行特异性分析,并与普通RT-PCR进行灵敏性比较。结果表明RT-LAMP法在30min就能够特异性鉴定出不同型别的IBV,且灵敏度是普通RT-PCR的107倍,反应结果可眼观判断。本研究建立的IBV RT-LAMP检测方法具备耗时短、灵敏度高、简单易行等特点,在实验室快速诊断中是一种检测IBV的有效方法。To develop a rapid,sensitive, and specific approach for the detection of infectious bronchitis virus（IBV）, reverse transcription-loop-mediated isothermal amplification（RT-LAMP） assay with four pairs of primers designed according to the conserved sequences of IBV N gene was established in the present study. With optimized reaction condition,the positive results of RT-LAMP assay were judged through visible green. Subsequently, the specificity was evaluated in detecting IBV, Newcastle disease virus, avian influenza virus and Mycoplasma synoviae of different strains. The viral RNA was amplified with Bst DNA polymerase within assay was spe here provided 30 min. The sensitivity was 107 times higher than that of conventional RT-PCR. RT-LAMP cific and simple to be operate with no need for special equipments. The approach established a promising method for the detection of IBV.

关 键 词：传染性支气管炎病毒 逆转录-环介导等温扩增 检测 

分 类 号：S852.659.6[农业科学—基础兽医学]