单管多重RT-PCR同时检测大豆种子中三种检疫性植物病毒  

Simultaneous detection of three quarantine plant viruses from soybean seeds by multiplex RT-PCR in single PCR tube

作　　者：易汪雪[1] 宋绍祎[1] 吴东妮[2] 代欢欢[3] 杨翠云[1] 于翠[1] 

机构地区：[1]上海出入境检验检疫局动植物与食品检验检疫技术中心,上海200135 [2]中国科学院上海生命科学研究院,上海生命科学信息中心,上海200031 [3]合肥师范学院,合肥230601

出　　处：《植物保护》2016年第5期113-117,共5页Plant Protection

基　　金：上海市科技兴农重点攻关项目[沪农科攻字(2012)第2-8号];上海出入境检验检疫局科研项目(HK005-2014)

摘　　要：我国大豆年进口量持续增长,快速、准确地检测进境大豆种子中可能携带的病原物是防止检疫性有害生物入境传播和扩散的有效手段。菜豆荚斑驳病毒(BPMV)、烟草环斑病毒(TRSV)和番茄环斑病毒(ToRSV)均是被大豆种子携带的检疫性有害生物。本研究建立了在单一PCR管中同时检测这3种检疫性植物病毒及大豆内源基因Bd-30K的多重RT-PCR方法。研究结果表明,所建立的多重RT-PCR方法具有较好的特异性和灵敏度,检测含BPMV、TRSV和ToRSV RNA的最低浓度分别为0.45、0.0093和0.004ng/μL,从大豆种子中同时检测3种病毒的最低RNA浓度为0.58ng/μL。The quantity of soybean seeds imported were continually increased in recent years.Rapid detection of seed-borne pathogens is essential to efficiently prevent pest invading and spreading.Bean pot mosaic virus（BPMV）,Tobacco ringspot virus（TRSV）and Tomato ringspot virus（ToRSV）are the quarantine pest which could be carried by the soybean seeds.We developed the multiplex RT-PCR approach for simultaneous detection of BPMV,TRSV,ToRSV and one endogenous gene Bd 30 K in the single PCR tube from soybean seeds.The results indicated that good specificity and sensitivity for simultaneous detection were obtained.The limit detection of RNA with BPMV,TRSV and ToRSV were 0.45 ng/μL,0.009 3 ng/μL and0.004 ng/μL,respectively.The limit detection of RNA with three viruses mixture was 0.58 ng/μL.

关 键 词：菜豆荚斑驳病毒 烟草环斑病毒 番茄环斑病毒 多重RT-PCR 同时检测 

分 类 号：S435.651[农业科学—农业昆虫与害虫防治] S432.41[农业科学—植物保护]