疫木中松材线虫的免疫学检测方法的研究  

Development and Application of an Immunoassay Method for Identification of Bursaphelenchus xylophilus in Pinewood

作　　者：白钢[1] 王玉嬿[2] 马洪周[1] 李海燕[2] 杨文博[1] 

机构地区：[1]南开大学生命科学学院,天津300071 [2]国家林业局森林病虫害防治总站,沈阳110034

出　　处：《林业科学》2005年第2期91-95,共5页Scientia Silvae Sinicae

基　　金：国家"十五"重点科技攻关课题"林业重大病虫害预警及控制技术研究"中的专题

摘　　要：以松材线虫的粉碎虫体免疫家兔制备抗血清 ,通过免疫印迹考察抗血清的特异性。利用固相松材线虫抗原和游离线虫抗原同线虫抗体相竞争 ,建立竞争型ELISA法用于疫木中松材线虫的分析。该方法可以直接检测出 10mg的木屑中 0 1μg微量的线虫蛋白 ,约 7条线虫的存在。利用该方法对 2 0个不同地区、树种及类型的松材样品进行检测 ,疫木中松材线虫的检出率为 10 0 % ,但拟松材线虫也呈现了一定的交叉反应。研究结果表明 ,以松材线虫抗原蛋白为指标的免疫学检测方法简便快速、灵敏度高 。Taking the specimens of Bursaphelenchus xylophilus (BX) and B. mucronatus (BM) fragments as research objects, an immunoassay method for nematode antigen analysis was developed in this paper. The rabbit antiserum against BX was prepared, and the antigen proteins were assayed by SDS_PAGE and western blot. Using BX fragments as solid_phase, the assay principle was based on competition between an analyte and nematode solid_phase antigen for anti_BX antibody. Then using HRP labeled goat anti_rabbit IgG as a tracer antibody, an enzyme_linked immunosorbent assay (ELISA) was developed. The method was specific to BX antigen harvested from different areas. Although the ELISA showed a little cross reactivity with BM, it could identify 0.1 μg nematode protein in 10 mg pinewood specimen. The result suggested that the ELISA method using nematode antigen as a detection objective showed a new potential for the quarantine inspection and forest pest control of BX.

关 键 词：检测方法 免疫学 ELISA法 拟松材线虫 免疫印迹 直接检测 交叉反应 研究结果 抗原蛋白 快速检疫 抗血清 特异性 检出率 灵敏度 竞争 家兔 虫体 

分 类 号：S852.4[农业科学—基础兽医学] S859.84[农业科学—兽医学]