世界主要储粮书虱环介导等温扩增试剂盒研发与应用

Development and application of loop-mediated isothermal amplification (LAMP) kit for main stored-grain booklice in the world

为利用分子生物学技术快速准确鉴定国内外主要储粮书虱,基于环介导等温扩增(loop-me‐diated isothermal amplification,LAMP)技术拟研发一款世界主要储粮书虱LAMP试剂盒,利用该LAMP试剂盒对10种储粮书虱的DNA粗提液进行测试,并结合DNA条形码技术对采自中国山东省和内蒙古自治区粮库的未知种类储粮书虱成虫样品进行鉴定。结果显示,本试验研发的LAMP试剂盒可准确鉴定10种储粮书虱;使用LAMP试剂盒和DNA条形码技术鉴定未知种储粮书虱样品的结果一致,即山东省储粮书虱样品为嗜虫书虱Liposcelis entomophila,内蒙古自治区储粮书虱样品为啮书虱L.corrodens,表明本试验研发的世界主要储粮书虱LAMP试剂盒可以快速、精准完成主要储粮书虱的鉴定,且操作便捷,可为海关口岸和基层仓储保护部门书虱鉴定工作提供技术支持。

In order to quickly and accurately identify main stored-grain booklice in the world by molecular identification technique, a loop-mediated isothermal amplification(LAMP) kit was developed.Based on crude DNA extract, the LAMP kit was tested against ten species of stored-grain booklice.Combined with DNA barcoding technique, this kit was used to identify the adult samples of unknown species of stored grain booklice collected from grain depots in Shandong Province and Inner Mongolia Autonomous Region, China. The results showed that the LAMP kit could accurately differentiated the ten species of stored-grain booklice. The results from the LAMP kit and DNA barcoding technique were consistent: the stored grain booklice sample from Shandong Province was Liposcelis entomophila, and the stored-grain booklice sample from Inner Mongolia Autonomous Region was L. corrodens, indicating that the LAMP kit developed in this study could quickly and accurately identify the main storedgrain booklice in the world, and the operation was simple. It could provide a technical support for the identification of stored grain booklice at ports of entry and in grass-roots storage protection departments.