应用等温环介导扩增方法检测蜜蜂残翅病毒的研究  

Diagnosis of the deformed wing cell virus( DWV ) by loop-mediated isothermal amplification

作　　者：庄明亮[1] 牛庆生[1] 陈东海[1] 王志[1] 李志勇[1] 梁勤[2] 

机构地区：[1]吉林省养蜂科学研究所,吉林132108 [2]福建农林大学,福州350002

出　　处：《环境昆虫学报》2016年第6期1192-1198,共7页Journal of Environmental Entomology

基　　金：国家蜂产业技术体系专项资金(CARS-45-KXJ7)

摘　　要：本文的目的在于建立用于临床检测残翅病病毒(Deformed wing virus,DWV)的等温环介导扩增技术(Loopmediated isothermal amplification,LAMP),为该疾病的预防和控制提供理论依据。在DWV基因保守序列设计4条引物,探究LAMP扩增的最优条件,并与常规的PCR(polymerase chain reaction)检测方法进行比较。建立的LAMP方法检测下限为0.89 pg,灵敏度比PCR高100倍而且特异性好。临床检测显示建立的LAMP方法可行、准确、方便、灵敏。针对DWV的LAMP建立的检测方法为养蜂生产第一线检测和预防DWV提供了技术支持,有一定的应用价值。The objective of this study is to establish a simple, fast and accurate method to deformed wing virus （DWV） in clinical by using loop-mediated isothermal amplification （LAMP）, which provides an experimental proof for controlling the disease. Four primers were designed based on six conserved regions of gene sequence on DWV, and were used for exploring the optimal LAMP amplification conditions. And the LAMP amplification result was compared conventional PCR （ polymerase chain reaction ） method. The established LAMP could detect as low as 0. 89 pg DNA, which was 100 times sensitive than PCR method. Clinical result showed LAMP could be used to detect DWV whatever in Italian bee （ Apis mellifera ligustica） or in Chinese bee （ A. cerana cerana ） , and the real positive detected ratio was 20% higher than PCR tested. The established LAMP was practical valuable for detecting and controlling the DWV in honeybee keeping.

关 键 词：残翅病毒 环介导等温扩增 检测 

分 类 号：Q963[生物学—昆虫学] S893.3[农业科学—特种经济动物饲养]