南方水稻黑条矮缩病毒检测方法的比较  

Comparison of detection methods for Southern rice black-streaked dwarf virus

作　　者：杜琳琳[1] 王跞姣 周彤[1] 兰莹[1] 范永坚[1] 周益军[1] 

机构地区：[1]江苏省农业科学院植物保护研究所,江苏省植物病毒病诊断检测技术中心,南京210014 [2]南京农业大学植物保护学院,南京210095

出　　处：《植物保护学报》2014年第3期352-359,共8页Journal of Plant Protection

基　　金：公益性行业(农业)科研专项(201303021);国家自然科学基金(3111130);江苏省农业科技自主创新资金(cx[12]1003);江苏省科技支撑计划(BE2012303);江苏省农业三新工程(SXGC[2013]359)

摘　　要：为明确南方水稻黑条矮缩病毒(Southern rice black-streaked dwarf virus,SRBSDV)检测方法的最佳适用范围,对其现有检测方法 Real time RT-PCR、RT-LAMP、RT-PCR的灵敏性及特异性进行了比较,并分析了依据SRBSDV单克隆抗体3F1建立的斑点免疫结合印迹(dot immunobinding assay,DIBA)方法对检测植物寄主和白背飞虱Sogatella furcifera Horvth的特异性。结果表明,灵敏性以Real time RT-PCR方法最高,其次为RT-LAMP方法,而普通RT-PCR方法相对较低。这3种方法均可特异性检测SRBSDV植物寄主和白背飞虱;DIBA方法可以满足SRBSDV和水稻黑条矮缩病毒(Rice black-streaked dwarf virus,RBSDV)植物寄主和白背飞虱大量样品的检测,但不能区分SRBSDV和RBSDV。Real time RT-PCR方法实现了短时间内对SRBSDV RNA拷贝数的相对定量;RT-LAMP方法全程恒温反应,无需热循环仪。To evaluate the optimal application range of detection methods for Southern rice black-streaked dwarf virus( SRBSDV),the sensitivities and specificities of existing SRBSDV detection methods,including Real-time RT-PCR,RT-LAMP and RT-PCR,were compared. And the specificities of dot immunobinding assay( DIBA) based on the monoclonal antibody 3F1 to SRBSDV used for detecting plants and white back planthopper,Sogatella furcifera,was analyzed. The results indicated that the Realtime RT-PCR is the most sensitive,followed by the RT-LAMP and the conventional RT-PCR. Real-time RT-PCR,RT-LAMP and RT-PCR were suitable for specifically detecting SRBSDV in plants and insect vectors. DIBA based on the monoclonal antibody 3F1 met the requirement for large-scale detection of SRBSDV and Rice black-streaked dwarf virus( RBSDV) in plants and insect vectors,but could not distinguish SRBSDV from RBSDV. Real-time RT-PCR could relatively quantify the copy number of SRBSDV RNA in a short time. RT-LAMP method is a reaction at constant temperature,which is feasible without a thermal cycler.

关 键 词：南方水稻黑条矮缩病毒 检测方法 比较 

分 类 号：S435.111.49[农业科学—农业昆虫与害虫防治]