3种核果类果树病毒多重PCR检测方法的建立与初步应用  

Establishment and application of multiplex PCR for diagnosing three quarantine viruses of stone

作　　者：赵世恒 王吉腾 李雪娇 高润蕾 云晓鹏[5] ZHAO Shiheng;WANG Jiteng;LI Xuejiao;GAO Runlei;YUN Xiaopeng(Beijing Tongzhou District Agricultural Product Quality Inspection Station,Beijing 100024,China;Rongcheng Entry-Exit Inspection and Quarantine Bureau, Rongcheng 264300, China;Plant Protection Station of Tongzhou District, Beijing 100024, China;Administrative Committee of Horinger Shengle Economic Development Zone, Horinger 011500 ,China;Plant Protection Institute, Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences,Hohhot 010031,China)

机构地区：[1]北京市通州区农产品质量检验检测站,北京100024 [2]荣成出入境检验检疫局,山东荣成264300 [3]北京市通州区植物保护站,北京100024 [4]呼和浩特市和林格尔县盛乐经济园区管委会,内蒙古和林格尔011500 [5]内蒙古农牧业科学院植物保护研究所,内蒙古呼和浩特010031

出　　处：《北方农业学报》2018年第3期37-41,共5页Journal of Northern Agriculture

基　　金：农业部呼和浩特作物有害生物科学观测试验站项目

摘　　要：根据李痘病毒(PPV)、李坏死环斑病毒(PNRSV)和李矮缩病毒(PDV)3种检疫性病毒的外壳蛋白基因保守序列设计特异性引物,在建立各种病毒单个RT-PCR技术的基础上,通过优化多重RT-PCR反应条件,初步建立了同步检测这3种病毒的多重RT-PCR检测体系。结果表明:3对引物特异性较强,分别可以扩增出172,675,945 bp的目的片段,该方法快速、灵敏、准确,为同时检测多种检疫性病毒提供了参考。The specific primers were designed according to the conserved sequences of coat protein genes of three quarantine viruses-Plum pox virus（PPV）, Lee dwarf virus（PDV） and Prunus necrotic ringspot virus（PNRSV） and the single detection of PPV, PDV and PNRSV was established by RT-PCR. Based on the above results a multiplex RT-PCR system of synchronously detecting those three virus was preliminarily established by optimization on of multiplex RT-PCR conditions.The experimental results showed that three pairs of the primers had strong specificity and could be used to amplify the 172,675,945 bp target DNAs. This method is rapid, sensitive and accurate, providing a reference for synchronously detecting several kinds of viruses in seedlings.

关 键 词：病毒检测 李痘病毒 李坏死环斑病毒 李矮缩病毒 多重RT-PCR 

分 类 号：S662[农业科学—果树学]