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以omp基因为靶标的3种检测方法对柑橘黄龙病菌检测效果比较

ISSN号：1004-874X
期刊名称：《广东农业科学》
时间：0
分类：S412[农业科学—植物保护]
作者机构：[1]广东省农科院植物保护研究所/广东省植物保护新技术重点实验室,广东广州510640, [2]南京农业大学植物保护学院,江苏南京210095
相关基金：广东省农业科技推广重大技术研究项目（201201127）;广东省现代农业产业技术体系建设专项（粤农[2009]380号）;广东省农业科学院院长基金（201305）;广东省科技创新人才出国培养计划项目（粤农科[2013]74号）

作者：程保平[1], 赵弘巍[2], 彭埃天[1], 陈霞[1], 宋晓兵[1], 凌金锋[1]

关键词：柑橘黄龙病, 常规PCR, LAMP, 实时荧光定量PCR, citrus huanglongbing, conventional PCR, LAMP, reahime quantitative PCR

中文摘要：

为了比较以omp基因为靶标的常规PCR、LAMP检测方法（Loop-mediated isothermal amplification）和实时荧光定量PCR方法在实际检测中对柑橘黄龙病的检测效果,比较了3种检测方法对柑橘黄龙病菌omp基因检测的灵敏度,并运用3种检测方法对采自广东省田间11个本地柑橘品种上疑似感染黄龙病的多个病样进行实际检测.结果显示：3种检测方法的灵敏度有所不同,依次为常规PCR＜LAMP检测方法=实时荧光定量PCR.3种检测方法对黄龙病田间样品的检出率依次为常规PCR＜LAMP检测方法＜实时荧光定量PCR.表明3种方法均可很好地检测黄龙病;其中常规PCR和实时荧光定量PCR适合有一定技术基础的农技人员使用,而后者具有最大的检测灵敏度,但成本较高;LAMP技术虽然是一种新技术,使用者较少,但该方法简单、快捷,且不需昂贵的精密仪器,具有很好的推广应用前景.

英文摘要：

Detection sensitivity and detection rate of conventional PCR, LAMP and real-time quantitative PCR were compared with the omp target gene. The results showed that LAMP had similar detection sensitivity with real-time quantitative PCR. And both of the two detection methods had higher detection sensitivity than that of the conventional PCR. Then total 35 suspected citrus samples from different cultivers with symptom of HLB in orchard of Guangdong were amplified and confirmed by conventional PCR, LAMP and real-time quantitative PCR. The results showed that LAMP had similar detection rate with real-time quantitative PCR. And both of the two detection methods had higher detection rate than that of the conventional PCR. This research indicated that conventional PCR could be used in large scale diagnosis of HLB disease, real-time quantitative PCR had the highest detection sensitivity, and LAMP was simple, fast and didn＇t rely on the expensive equipments, had good application prospect.