食源性致病菌焦磷酸通用引物测序方法的建立  

The Study of Pyrosequencing Method by the Universal Primers for Food-borne Pathogenic Bacteria

作　　者：赵新 王永 兰青阔 陈锐 朱珠 余景会 李欧静 郭永泽 

机构地区：[1]天津市农业质量标准与检测技术研究所,天津300381

出　　处：《食品研究与开发》2013年第1期97-100,共4页Food Research and Development

基　　金：科技部国际合作项目(2006DFA32380);天津市农业科学院院长基金项目(08024)

摘　　要：建立一种利用焦磷酸测序技术检测和鉴定沙门氏菌(Salmonellla)、单核细胞增生李斯特菌(Listeria monocytogenes)、金黄色葡萄球菌(Staphylococcus aureus)和志贺氏菌(Shigella)的方法。通过对4种菌16SrRNA的保守序列可变区的同源性分析,设计出一套适合焦磷酸测序的扩增引物和测序引物,通过可变区位点差异,利用一套通用引物同时鉴别4种食源性致病菌,测序结果与数据库比对判断细菌种属。结果表明:建立的检测方法可在4 h内完成对4种菌的鉴定,测序鉴定结果与国标的传统生化方法完全一致,焦磷酸测序技术可用于食源性致病菌的检测和鉴定。The method of pyrosequencing technology for identification of Salmonellla spp, Listeria monocytogenes, Staphylococcus aureus and Shigella cereus was established. According to nucleotide homology analysis of 16S rRNA of 4 food-borne pathogenic bacteria,we designed one set of primer,including one universal PCR primers and one pyrosequencing sequencing primer. The 4 pathogenic bacteria were distinguished on the basis of pyrosequencing signal peaks. The result showed that the identification of four-kinds of food-borne pathogenic bacteria was completed in 4 h, the results of identification of bacteria were consistent with the result of GB/T method. The pyrosequencing method can use for detection and identification of food- borne pathogenic bacteria.

关 键 词：食源性致病菌 焦磷酸测序技术 通用引物 16SrRNA 

分 类 号：TS207.4[轻工技术与工程—食品科学]