枣疯病植原体LAMP可视化检测方法的建立  

Development of a Loop-mediated Isothermal Amplification Assay for Visual Detection of Jujube Witches' Broom Phytoplasma

作　　者：韩剑[1] 罗明[1] 何贵伦 张祥林[2] 项雪峰[3] 

机构地区：[1]新疆农业大学农学院,乌鲁木齐830052 [2]新疆出入境检验检疫局,乌鲁木齐830063 [3]新疆阿克苏地区农业检验监测中心,新疆阿克苏843000

出　　处：《西北农业学报》2015年第6期125-131,共7页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：新疆维吾尔自治区高校科研计划(XJEDU2010S15)

摘　　要：根据枣疯病植原体16SrDNA基因保守序列,设计出4条特异性环介导等温扩增(LAMP)引物。通过对反应条件的优化,建立一种适用于枣疯病植原体的可视化LAMP检测技术,对优化后的方法进行特异性、灵敏度评价。结果显示,该方法能够特异性地检测出枣疯病植原体,且LAMP产物的特异性通过MaeⅡ酶切得到验证;灵敏度较常规PCR高出100倍;并可通过肉眼观察反应液颜色变化,直接判定结果。建立的枣疯病植原体LAMP可视化检测方法适用于基层及现场快速检测,为枣疯病的快速检测提供了新方法。The objective of this study is to establish a quick and accurate detection technique of jujube witches＇ broom phytoplasma by loop-mediated isothermal amplification（LAMP）.A set of four LAMP primers specific for jujube witches＇ broom phytoplasma were designed according to its conserved region of 16 SrDNA gene.Based on optimization of reaction conditions,LAMP detection method was developed which was treated with specific techniques and was evaluated.The result showed that the loop-mediated isothermal amplification method has specificity amplification action only for jujube witches＇ broom phytoplasma,and the specificity of LAMP products were confirmed by digestion with MaeⅡ restriction enzyme.The sensitivity for the detection of jujube witches＇ broom phytoplasma was 100 times higher than that of the conventional PCR method.Detection results can be judged by visual color change and not require complex equipment.The method was very suitable for detection in the field condition that would provided a novel rapid detection approach for jujube witches＇ broom.

关 键 词：枣疯病 植原体 环介导等温扩增 可视化检测 

分 类 号：S436.65[农业科学—农业昆虫与害虫防治]