基于环介导等温扩增技术快速检测美澳型核果褐腐病菌  

Rapid detection of Monilinia fructicola using a loop-mediated isothermal amplification assay

作　　者：王颖[1] 徐倩 孙旻旻[1] 李艳华[1] 周培[1] 曹志强[1] 李彬[3] 郑小波[2] 

机构地区：[1]无锡出入境检验检疫局,江苏无锡214000 [2]南京农业大学植保学院 [3]江苏出入境检验检疫局

出　　处：《植物检疫》2017年第5期32-38,共7页Plant Quarantine

基　　金：国家质量监督检验检疫总局科技计划项目(2015IK157;2016IK145);无锡出入境检验检疫局科技计划项目(2016WX01)

摘　　要：本文建立了美澳型核果褐腐病菌(Monilinia fructicola)环介导等温扩增技术,并利用此技术开展了M.fructicola的检测和鉴定。采用M.fructicola SCAR分子标记的MO368特异片段为靶标序列,设计并筛选出M.fructicola的特异性LAMP引物,建立了该病菌快速诊断方法。试验结果表明,仅M.fructicola的菌株DNA扩增后呈天蓝色的阳性反应,而在其他真菌的供试菌株中均呈紫色的阴性反应。检测灵敏度可达到100 pg/μL。利用该方法对无锡机场截获的7份疑似李子褐腐病样本进行检测,结果有3份样品呈LAMP阳性反应。本文建立的M.fructicola LAMP检测方法具有灵敏度高、特异性好,简便易行等优点,为美澳型核果褐腐病菌的快速检测提供了一种新技术。This study reports the development of a loop-mediated isothermal amplification（LAMP） assay targeting the MO368 SCAR for visual detection of Monilinia fructicola.By comparing different sequences of M. fructicola with those of relative species,we selected MO368 SCAR as the target gene,designed and selected a set of specific and sensitive LAMP primers to detect M. fructicola. A positive color（sky blue） was only observed in the presence of M. fructicola whereas all other isolates showed purple color.The detection limit of the LAMP assay for M. fructicola was 100 pg/μL.M. fructicola was detected from 3 among 7 suspected disease samples intercepted at Wuxi airport in China.This method provides a new technology for the diagnosis and quarantine of M. fructicola.

关 键 词：美澳型核果褐腐病菌 环介导等温扩增 检测 

分 类 号：S41-30[农业科学—植物保护]