摘要
介绍一种便捷、灵敏而又特异的用于检测治病细菌小肠结肠炎耶尔森氏菌(Yersinia enterocolitica)的环介导恒温扩增基因检测技术(loop-mediated isothermal amplification,LAMP),该技术分别使用特异对应于16S-23S rDNA间区(16S-23S rDNA internal transcribed spacer,ITS)靶序列中6个基因区段的3对引物,在Bst聚合酶的作用下对靶序列进行恒温扩增,整个检测反应只需1 h.利用这种技术检测小肠结肠炎耶尔森氏菌基因,阳性结果可通过直接观察反应液中有无焦磷酸镁白色沉淀,而无需经电泳检测.试验结果表明,LAMP对Y.enterocolitica的检测灵敏度可达到8 CFU/反应.
Yersinia enterocolitica（Y.enterocolitica） is an important human enteroinvasive pathogen.Here we developed a loop-mediated isothermal amplification（LAMP） method to detect Y.enterocolitica in food.The six primers of the LAMP targeted at 16S 23S rDNA internal transcribed spacer（ITS） are high specific to differiate Y.enterocolitica from the other Yersinia species and the other bacteria species.This method is more sensitive and efficient than that of PCR,the organism can be identified within 1 hour at a minimal amount of bacteria with 8 CFU per reaction.
出处
《南开大学学报（自然科学版）》
CAS
CSCD
北大核心
2010年第2期8-14,共7页
Acta Scientiarum Naturalium Universitatis Nankaiensis
基金
天津出入境检验检疫局项目(tk028-2008)
关键词
耶尔森氏菌
快速检测
16S-23S
RDNA间区
环介导恒温扩增
Yersinia enterocolitica
rapid detection
16S-23S rDNA internal transcribed spacer（ITS）
loopmediated isothermal amplification（LAMP）
