免疫磁性捕获ELISA技术在松材线虫检测中的应用  

Development and Application of Immunomagnetic Separation ELISA for Identification of Bursaphelenchus xylophilus in Pinewood

作　　者：曹宇[1] 李海燕[2] 马洪周[1] 胡加付[2] 杨文博[1] 王玉嬿[2] 白钢[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,辽宁沈阳110034

出　　处：《林业科学研究》2005年第5期585-589,共5页Forest Research

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

摘　　要：本研究采用种子聚合法合成聚苯乙烯磁性微球,并以兔抗松材线虫IgG致敏,制备出能特异性地捕获松材线虫蛋白抗原的免疫磁性微球。以生物素标记抗体为示踪抗体,并结合酶标亲和素检测系统,用于疫木样品的分析。利用该方法对采自不同地区的3种松树,其中松材线虫病木17株、拟松材线虫病木5株和健木3株进行检测。实验结果表明,虽然拟松材线虫病木也呈现了一定的交叉反应,但疫木中松材线虫总检出率为94.1%,灵敏度达到0.1μg.mL-1线虫蛋白抗原。研究表明免疫磁性捕获ELISA技术可直接捕获木屑中的微量线虫抗原,具有简便、快速、准确等优点,是一种实用的松材线虫的快速检疫方法。Using dispersion polymerization, polystyrene magnetic micropsheres as carriers were prepared. By coating with rabbit anti-Bursaphelenchus xylophilus （BX） IgG, the immunomagnetic microspheres （IMMS） could specifically capture nematode antigen on pinewood samples. Taking biotinylated Anti-BX rabbit IgG as a tracer antibody, together with horseradish peroxidase labeled avidin, an enzyme-linked immuosorbent assay （ELISA） on IMMS was developed. Three kinds of pine trees harvested from different areas were detected by this method. Although there were a few crossed reactivity between Bursaphelenchus mucronatus antigen with Anti-BX IgG, the relevance ratio of BX on infected pine trees was 94. 1% and the detection limit could be 0.1 μg · mL^-1 nematode antigen. The result suggested that immunomagnetic separation enzymelinked immuosorbent assay （IMS-ELISA） could directly capture trace nematode antigen on pinewood homogenizations. The method showed practicability of the quick quarantine inspection for its speediness, convenience and accuracy.

关 键 词：免疫磁性微球 免疫磁性捕获 ELISA 松材线虫 

分 类 号：S763[农业科学—森林保护学]