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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。
In this study, polystyrene magnetic microspheres were synthesized by seed polymerization and sensitized with rabbit anti-pine nematode IgG to prepare immunomagnetic microspheres that can specifically capture the antigen of pine wood nematode protein. The biotin-labeled antibody was used as the tracer antibody, and the enzyme-linked avidin detection system was used for the analysis of the diseased wood samples. Using this method, three kinds of pine trees from different regions, including 17 pine wood nematode, 5 pine wood nematode and 3 wood from Jianmu were tested. The experimental results showed that, although the pine wood nematode disease also showed a certain degree of cross-reaction, but the overall detection rate of pine wood nematode was 94.1%, the sensitivity of 0.1μg.mL-1 nematode protein antigen. The results show that the immunomagnetic capture ELISA technique can directly capture trace nematode antigens in sawdust, which is simple, rapid and accurate. It is a practical quarantine method for pine wood nematode.