3种番茄斑萎病毒属病毒多重RT-PCR检测方法的建立  

A method of synchronously detecting three kinds of Tospovirus viruses by multiplex RT-PCR

作　　者：代欢欢[1,2] 陈舜胜[1] 杨翠云[2] 于翠[2] 

机构地区：[1]上海海洋大学食品学院,上海201306 [2]上海出入境检验检疫局,上海200135

出　　处：《上海农业学报》2012年第3期32-36,共5页Acta Agriculturae Shanghai

基　　金：国家质检总局科技项目(2011IK172)资助

摘　　要：利用LaserGene软件对Genbank数据库中已登陆的番茄斑萎病毒(TSWV)、凤仙花坏死斑病毒(INSV)和鸢尾黄斑病毒(IYSV)全基因组序列进行比对,选择最保守的区域,设计3种病毒的特异性引物,通过逐步调整优化PCR的反应条件,建立了多重RT-PCR检测TSWV、INSV和IYSV的技术体系。结果表明:可从约16μg的带毒烟草叶片中同时检测到3种病毒,具有较高的特异性和准确性。该方法可以同步、快速、特异地检测出受侵染寄主植物中的TSWV、INSV和IYSV。The most conserved regions were selected via aligning the total genomic sequences of to- mato spotted wilt virus（TSWV）, Impatiens necrotic spot virus（INSV）and iris yellow spot virus（IYSV） in the Genebank by using the LaserGene software, the viruses＇specific primers were designed, and a method of detecting TSWV, INSV and IYSV by multiplex RT-PCR was established through progres- sively adjusting and optimizing the PCR conditions. The results showed that the method could synchro- nously detect TSWV, INSV and IYSV from an about-16 μg infected tobacco leaf and had relatively high specificity and accuracy. The method could detect TSWV, INSV and IYSV of infected host plants synchronously, rapidly and specifically.

关 键 词：病毒检测 番茄斑萎病毒 凤仙花坏死斑病毒 鸢尾黄斑病毒 多重RT-PCR 

分 类 号：S436.412[农业科学—农业昆虫与害虫防治]