基于TC-RT-PCR方法的几种烟草主要病毒病的多重检测体系的构建  

Multiple Detection System Establishment of Several Main Virus Diseases Based on TC-RT-PCR

作　　者：任锡毅 刘永翔[1,2] 黄永会[1,2] 刘作易[2,3] 

机构地区：[1]贵州省生物技术研究所,贵州贵阳550006 [2]贵州省农业生物技术重点实验室,贵州贵阳550006 [3]贵州省农业科学院,贵州贵阳550006

出　　处：《贵州农业科学》2015年第9期15-19,22,共6页Guizhou Agricultural Sciences

基　　金：中国烟草总公司贵州公司科技项目"贵州烟草主要病毒病调查与快速检测研究"(201111)

摘　　要：为避开病毒病检测中分子生物学方法检测时RNA提取这一复杂费时费力的过程,且能在同一个反应中同时检测多种病毒病,使大量病毒病样品和多种病毒病复合侵染的检测更简单、快捷。对我国常见的3种烟草病毒的外壳蛋白基因部分序列设计引物,通过优化反应条件,摸索扩增参数构建多重TC-RTPCR检测体系。结果表明:首次成功构建能同时检测TMV、CMV和PVY的多重TC-RT-PCR检测体系。该技术无需RNA的提取,包括叶片研磨、捕捉、反转录、PCR扩增和电泳检测等步骤,简单快捷、省时省力,具有高效性、系统性和经济简便性等特点。The multiple TC-RT-PCR detection system was established by amplification parameters through the optimized reaction condition after the primers for partial sequence of CP gene of three common tobacco viruses in China were designed to avoid the complex,waste time and energy process of RNA extraction in virus detection,detect several viruses in the same reaction simultaneously and probe a detection method with simplicity and rapidness for a large number of virus samples and multiplex infection of multiple viruses.Results：the multiple TC-RT-PCR detection system to simultaneously detect TMV,CMV and PVY is successfully established for the first time.The multiple TC-RT-PCR detection system has the advantages of simplicity,rapidness,saving time and labor,high efficiency,systematicness and economy because it does not need the process of RNA extraction including leaf grind,capture,reverse transcription,PCR amplification and electrophoresis detection.

关 键 词：烟草病毒 多重TC-RT-PCR 检测 

分 类 号：S431.16[农业科学—农业昆虫与害虫防治]