玉米细菌性枯萎病菌TaqMan探针实时荧光PCR检测方法的建立  

Detection and identification of Pantoea stewartii stewartii using TaqMan probe

作　　者：漆艳香[1] 朱水芳[2] 赵文军[2] 肖启明[1] 廖晓兰[1] 

机构地区：[1]湖南农业大学 [2]国家质检总局动植物检疫实验所,北京100029

出　　处：《植物保护学报》2004年第1期51-56,共6页Journal of Plant Protection

基　　金：国家质检总局植物病原检测专项(Z2000-3-182)

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。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

关 键 词：玉米 细菌性枯萎病菌 TAQMAN探针 实时荧光PCR 检测方法 细菌性枯萎病 

分 类 号：S435.13[农业科学—农业昆虫与害虫防治]