---

期 刊 ：植物保护学报 2008年

关键词：灰飞虱；水稻条纹病毒；免疫检测；RT-PCR检测；

Keywords:Laodelphax striatellus, Rice stripe virus, immune-detection, RT-PCR assay,

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 Falln (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。

Abstract:In order to estimate serviceable range of the detection methods of Rice stripe virus (RSV) in small brown plant hopper,Laodelphax striatellus Falln (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.

---