摘要
为评价灰飞虱体内水稻条纹病毒检测方法的适用范围,利用已经制备的水稻条纹病毒(Rice stripe virus,RSV)的多克隆抗体SV21和单克隆抗体3B9,采用多孔板间接ELISA、DIBA和Western blotting等三种方法进行单头灰飞虱Laodelphax striatellus Fallén(SBPH)体内RSV检测。结果表明,检测灵敏度以多孔板间接ELISA最高,其次为DIBA,Western blotting最低;用单克隆抗体3B9检测灵敏度高于多克隆抗体SV21。RT-PCR、IC-RT-PCR和DB-RT-PCR三种方法中,RT-PCR对单头灰飞虱稀释到400倍可检测到病毒;IC-RT-PCR在多克隆抗体SV21稀释浓度大于500倍时检测不出RSV,单克隆抗体3B9稀释浓度大于800倍时检测不到RSV;DB-RT-PCR检测结果显示,单头灰飞虱在稀释400倍后均无阳性反应。
In order to estimate serviceable range of the detection methods of Rice stripe virus （RSV） in small brown plant hopper, Laodelphax striateUus Fallen （SBPH）, polyclonal antibody SV21 and monoclonal antibody 3B9 against RSV were used. RSV in a single SBPH could be detected with the ELISA, DIBA and Western blotting methods successfully. The monoclonal antibody 3B9 was more sensitive than polyclonal antibody SV21 and polywell-plate indirect-ELISA with polyclonal antibody or monoclonal antibody as testing antibody had a higher specificity and sensitivity than DIBA and Western blotting. Among the three molecular detection methods, RSV could be detected with 1：400 dilution of extraction of a single SBPH using RT-PCR, however, RSV could not be detected using IC-RT-PCR if the dilution of polyclonal antibody SV21 was less than 1： 500 or the dilution of monoclonal antibody 3B9 was less than 1：800. The results of DB-RT-PCR showed that RSV could not be detected with less than 1：400 dilution of a single male or female SBPH.
出处
《植物保护学报》
CAS
CSCD
北大核心
2008年第5期410-414,共5页
Journal of Plant Protection
基金
江苏省重大科技攻关项目(BE2005301)
国家支撑计划(2006BAD02A16、2006BAD08A04)
江苏省自然科学基金(BK2006164)
农业部行业专项(nyhyzx07-51)
江苏省333人才培养工程资助项目
国家自然科学基金(30300230)
