【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。更多还原

【Abstract】 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.更多还原