鸭Ⅰ型肝炎病毒RT-LAMP可视化检测方法的建立  

Development of a reverse transcription loop-mediated isothermal amplification assay for visual detection of duck hepatitis virus type Ⅰ

作　　者：谢丽基[1] 谢芝勋[1] 刘加波[1] 庞耀珊[1] 邓显文[1] 谢志勤[1] 范晴[1] 

机构地区：[1]广西壮族自治区兽医研究所,广西南宁530001

出　　处：《中国预防兽医学报》2012年第2期112-115,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家百千万人才工程人选专项资金项目(945200603);广西科技攻关项目(桂科攻10100014-5);桂科专项11-3

摘　　要：为建立一种快速检测鸭Ⅰ型肝炎病毒(DHV I)的方法,本研究根据基因库中DHV Ⅰ基因的保守序列,设计特异性环介导等温扩增(LAMP)引物,建立了DHV I的RT-LAMP可视化检测方法。该方法的敏感性可达10fg,高于常规PCR方法 100倍;全部反应可在1 h内完成;可以通过肉眼观察颜色直接判定结果;对其它鸭常见病原体的检测结果均为阴性。实验结果表明建立的RT-LAMP方法简便、快速、灵敏、特异,可用于DHV Ⅰ感染的快速检测。In this study, a reverse transcription loop-mediated isothermal amplification （RT-LAMP） assay was developed lbl detection of duck hepatitis virus type I （DHV I） with six primers designed according to the sequences of DHV I in GcnBank. Under the optimized reaction conditions, the test results showed that the RT-LAMP method was spcefic and no cross-rcac｜ivity with the other pathogen of duck. The detection limit of the RT-LAMP was 10 fg of virus RNA, which was higher than the routine PCR. The amplification was completed within one hour, and the detection result could be judged even by naked eyes. These results demonstrated that the RT-LAMP assay is a simple and specific method for rapid detection of DHV I in clinical samples.

关 键 词：鸭Ⅰ型肝炎病毒 环介导等温扩增 可视化检测 

分 类 号：S852.65[农业科学—基础兽医学]