小麦矮缩病毒NASH快速检测方法的建立及应用  

Development and application of nucleic acid spot hybridization(NASH)assay for rapid detection of Wheat dwarf virus

作　　者：金文[1] 张金良 刘艳[1] 王锡锋[1] 

机构地区：[1]中国农业科学院植物保护研究所,植物病虫害生物学国家重点实验室,北京100193 [2]北京市植物保护站,北京100029

出　　处：《植物保护》2015年第3期100-103,共4页Plant Protection

基　　金：国家自然科学基金(31171820);北京市粮食高产创建项目(PXM2013-036203-000022);粮食作物产业技术体系北京市创新团队项目

摘　　要：以非放射性物质地高辛为标记物,采用PCR法制备了特异性强、灵敏度高的DNA探针。通过优化反应体系,建立了小麦矮缩病毒(Wheat dwarf virus,WDV)的核酸斑点杂交(nucleic acid spot hybridization,NASH)快速检测技术体系。该方法诊断准确率高,操作简单,周期短,整个检测过程仅需5h左右。利用建立的NASH技术开展WDV流行学调查,发现近年来WDV在我国陕西韩城、山西太原和河北石家庄等地区点片发生,没有大面积暴发成灾。In this study, a nucleic acid spot hybridization （NASH） method was developed to detect Wheat dwarf virus （WDV）. Based on digoxigenin （non-radioactive）-labeled strategy, a DNA probe with well-characterized specificity and sensitivity was prepared by PCR. The reaction system was optimized for highly accurate diagnosis and simple operation. The whole process of assay was rapid and efficient, which could be completed in 5 h. The established system was applied to epidemiological survey of WDV. The results indicated that no large-scale outburst of WDV has occurred in China in recent years, and it only happened sporadically in local areas, including Hancheng （Shaanxi Province）, Taiyuan （Shanxi Province） and Shijiazhuang （Hebei Province）.

关 键 词：小麦矮缩病毒 地高辛 核酸斑点杂交(NASH) 快速检测 

分 类 号：S435.121.5[农业科学—农业昆虫与害虫防治]