实时荧光RT-PCR检测辣椒轻斑驳病毒的初步研究  

A preliminary study on detection of Pepper mild mottle virus by real-time fluorescent RT-PCR

作　　者：郭京泽[1] 李兴红[2] 廖芳[1] 张文明[2] 刘鹏[1] 刘跃庭[1] 

机构地区：[1]天津出入境检验检疫局,天津300456 [2]北京农林科学院植物保护环境保护研究所,北京100097

出　　处：《植物保护》2008年第2期117-120,共4页Plant Protection

基　　金：北京市自然科学基金项目(6052010);天津检验检疫局课题(TK001-2007)

摘　　要：本研究选取辣椒轻斑驳病毒PMMoV、烟草花叶病毒TMV、黄瓜花叶病毒CMV和马铃薯Y病毒PVY作为试验材料,根据PMMoV衣壳蛋白(CP)RNA的序列特异性位点,设计出Taqman荧光探针及其引物,采用实时荧光RT-PCR技术对PMMoV进行快速检测,同时与ELISA方法进行了灵敏度比较,结果表明:该方法具有较高的灵敏度及较强的特异性,PMMoV检测结果为阳性,TMV、CMV和PVY均无荧光信号,为阴性,灵敏度是传统的ELISA方法的100倍,而且大大缩短了检测时间。该方法快速、准确、灵敏、简便、安全,具有实际应用价值,适用于植物病毒病害的快速检测。As the pathogen of a main disease, Pepper mild mottle virus （PMMoV） severely endangers its natural host the sweet pepper. This study chose PMMoV, TMV, CMV and PVY as test materials, and special Taqman flurescent probes and primers for PMMoV were designed based on the sequence of coat protein gene. Based on realtime fluorescent RT-PCR, a credible, sensitive, safe and special detection method for PMMoV was successfully established. Compared with the DAS-ELISA method, the sensitivity of this detection assay was increased by one hundred times. Moreover, this real-time fluorescent RT-PCR greatly shortened the detection time. The establishment of this detection method would strongly promote the quarantine of plant seed-borne virus diseases.

关 键 词：辣椒轻斑驳病毒 实时荧光RT-PCR 双抗夹心酶联免疫吸附法 

分 类 号：S436.418.12[农业科学—农业昆虫与害虫防治]