焦磷酸测序技术在4种食源性致病菌快速检测中的应用

为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种菌毒力靶基因特异性序列的同源性分析,设计出适合焦磷酸测序特异性扩增和测序的引物,摸索出最佳的焦磷酸测序反应体系和反应程序,并建立了4种食源性致病菌焦磷酸测序快速检测方法。此方法以传统国标法的前增菌为前提,而省去了后期生化验证的繁琐过程,大大缩短了检测时间,并且其准确性与传统生化验证完全一致,灵敏度可达10 CFU/mL。结果表明,作者建立的焦磷酸测序检测方法具有高效、快速、精准及操作简便等特点,为食源性致病菌的快速鉴定开辟了广阔的前景。

Study and Application on Detection of Four Kinds of Food-Borne Pathogenic Bacteria by Pyrosequencing Technology

The aim of this study was to establish a detection method for identification of food borne pathogenic bacteria by using pyrosequencing technology.For this,one pair of polymerase chain reaction(PCR) primers and one pyrosequencing sequencing primer of Salmonellla spp,Listeria monocytogenes,Staphylococcus aureus and Shigella cereus was designed,then the pyrosequencing reaction system and conditions was optimized.the sensitivity of this method was arrived at 10 CFU/mL and has been proven to be highly efficiency,rapid,exact and easy handling.The results provide an informative supplement to fast identified food-borne pathogenic bacteria.