鳞球茎茎线虫LAMP检测方法的建立

Development of LAMP assay for rapid detection of Ditylenchus dipsaci

鳞球茎茎线虫(Ditylenchus dipsaci)能引起多种植物毁灭性病害,是世界公认的危险性线虫,被列入我国检疫性有害生物名录。本研究通过比较GenBank数据库中茎属线虫相关序列,以鳞球茎茎线虫rDNA-ITS序列为靶标,设计鳞球茎茎线虫LAMP特异性引物,建立了可视化LAMP检测体系。结果表明,本研究建立的LAMP检测方法可在1 h内对鳞球茎茎线虫进行检测,供试其他22种植物线虫均未出现扩增,仅鳞球茎茎线虫出现扩增,具有良好的特异性;检测灵敏度可达1/4000条鳞球茎茎线虫;检测结果可以直接用肉眼判断。因此,本方法具有反应时间短、灵敏度高、设备要求低的优点,适于基层实验室开展鳞球茎茎线虫检测。

Ditylenchus dipsaci is a globally recognized dangerous nematode that can cause various destructive plant diseases and is listed as a quarantine pest in China.This study compared the sequences related to Ditylenchus in the GenBank database,and chose to use the rDNA-ITS sequence of D.dipsaci as the target.Then LAMP specific primers were designed and a visual LAMP detection system was established.The results showed that the LAMP detection method can detect D.dipsaci within 1 hour.Meanwhile,only D.dipsaci showed amplification,and the other 22 plant nematodes didn't amplify.The detection sensitivity can reach 1/4000 scale of D.dipsaci.Furthermore,the detection results can be directly judged with the naked eye.Therefore,this method has the advantages of short reaction time,high sensitivity,and low equipment requirements,making it suitable for grassroots laboratories to carry out the detection of D.dipsaci.