柑橘黄龙病菌RAA检测技术研究与应用

Research and Application of RAA Detection Technology for Candidatus Liberibacter

以柑橘黄龙病亚洲韧皮杆菌保守基因序列为靶基因设计特异性引物,经引物筛选和反应条件的优化,建立黄龙病菌的重组酶等温扩增的快速检测方法。该方法在39℃反应温度下20min即可实现靶标基因片段的有效扩增。特异性分析结果表明RAA方法检测柑橘黄龙病病菌与其他常见致病菌菌株间不存在交叉反应,结果表明,RAA方法操作简单、费用低廉、特异性强、灵敏度高、快速高效、操作简单、检测结果直观、对仪器设备要求低等特点,是一项具有广泛前景的方法,为检验黄龙病菌的检测提供新的发展方向。

Specific primers are designed using the conserved gene sequences of Candidatus Liberibacter asiaticus as the target gene.After primer screening and optimization of reaction conditions,a rapid detection method of Recombinase Aided Amplification,RAA is established.This method enables efficient amplification of target gene fragments for 20 min at a reaction temperature of 39℃.The results of the specific analysis shows that the RAA method detected no cross-reaction between the citrus yellow dragon disease bacteria and other common pathogenic bacteria.The results show that the RAA method is characterized by simple operation,low cost,strong specificity,high sensitivity,fast and high efficiency,simple operation,intuitive test results and low instrument and equipment requirements,which is a widely promising method and provides a new development direction for the detection of yellow shoot.