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当前位置: 首页 > 文章 > 玉米细菌性枯萎病菌TaqMan探针实时荧光PCR检测方法的建立 植物保护学报 2004,31 (1) 51-56
Position: Home > Articles > Detection and identification of Pantoea stewartii stewartii using TaqMan probe Journal of Plant Protection 2004,31 (1) 51-56

玉米细菌性枯萎病菌TaqMan探针实时荧光PCR检测方法的建立

作  者：
漆艳香;朱水芳;赵文军;肖启明;廖晓兰
单  位：
湖南农业大学;国家质检总局动植物检疫实验所
关键词：
TaqMan探针;实时荧光PCR;玉米细菌性枯萎病菌;检测;鉴定
摘  要：
成功建立了玉米细菌性枯萎病菌快速检测鉴定的实时荧光PCR方法。该方法根据细菌16S rDNA序列的特异性,设计出对玉米细菌性枯萎病菌具有稳定性点突变特异性探针,并对10种细菌菌株和5种植原体进行了实时荧光PCR。结果表明,只有玉米细菌性枯萎病菌产生荧光信号,而其它参考菌不产生荧光信号,检测的绝对灵敏度是14.2 fg/μl质粒DNA,比常规的PCR电泳检测高约100倍。整个检测过程只需2h,完全闭管,降低了污染的机会,无须PCR后处理。
译  名：
Detection and identification of Pantoea stewartii stewartii using TaqMan probe
作  者：
Qi Yanxiang Zhu Shuifang Zhao Wenjun Xiao Qiming Liao Xiaolan (College of Plant Protection, Hunan Agriculture University, Changsha 410128, China; Animal and Plant Quarantine Institute, State Administration of Entry-Exit Inspection and Quarantine of China, Beijing 100029, China; Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou 571737,China)
关键词：
TaqMan probe;real-time fluorescent PCR;Pantoea stewartii stewartii;detection ;identification
摘  要：
Pantoea stewartii stewartii is a very important quarantine bacterium on the Av1 list of China,which has not been reported in China yet.The biological methods and ELISA were used mainly for detection of P.s.stewartii.A group-specific probe was designed based on 16S rDNA of P.s.stewartii and it was used to detect 3 species of Pantoea,3 species of Erwinia,4 species of other bacteria and 5 species of phytoplasma.The results show that no signal was detected for other bacteria and phytoplasma except P.s.stewartii.The sensitive level of detection of probe was 14.2 fg/μl plasmid DNA and 100 times of that in PCR gel electrophoresis detection.Little contamination will occur because the whole detection process is finished in the contained tubes,so the method is more specific than normal PCR.TaqMan probe designed in the studv can detect P.s.stewartii immediatelv and directly

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