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诊断松萎蔫病的传统方法是从木质组织中分离松材线虫,然后用显微镜进行形态学鉴定。基于DNA分子检测的方法尽管很灵敏,不需要固定发育阶段的松材线虫,但是仍然需要用Baer-mann漏斗法分离线虫,并且需要昂贵的仪器。笔者研发了利用环介导恒温扩增法检测松材线虫的存在,该方法包括:(1)从木块中提取DNA;(2)DNA扩增;(3)通过反应溶液的颜色做出诊断。该方法仅使用培养箱且整个过程只需要90 min,比以往方法更加简便和快速。
The traditional method for the diagnosis of pine wilt disease is to separate the pine wood nematode from the woody tissue and then use a microscope for morphological identification. The method based on DNA molecular detection, though very sensitive, does not require immobilized stages of pine wood nematode, but nematodes still need to be isolated using the Baer-mann funnel method and require expensive instrumentation. The authors have developed the use of ring-mediated isothermal amplification for the detection of Bursaphelenchus xylophilus, comprising: (1) DNA extraction from wood chips; (2) DNA amplification; (3) diagnosis by color of the reaction solution . This method only uses the incubator and the entire process takes only 90 min, which is easier and faster than before.