机构地区: 河北农业大学
出 处: 《农产品加工(下)》 2015年第9期49-52,共4页
摘 要: 利用环介导等温扩增技术(LAMP)检测牛肉中大肠杆菌O157。以O157∶H7的rfb E基因序列(Genbank S83460)为靶序列设计了5条引物,通过可视白色沉淀和电泳观察结果。19株食源性致病菌被应用于LAMP方法检测的特异性试验,其中显示阳性结果的为3株大肠杆菌O157,剩余16株菌结果显示均为阴性。LAMP检测大肠杆菌O157∶H7的灵敏度为1.0 CFU/m L,人工污染牛肉的检出限为1.4 CFU/g。LAMP技术作为一种基因检测技术,因其灵敏度高、特异性强以及操作简单等特点,将为食源性致病菌的检测搭建一个新的技术平台,尤其在大肠杆菌O157∶H7的快速检测上存在巨大潜力。 The loop-mediated isothermal amplification (LAMP) technique is used to detect Escherichia coli O157 in beef. Five primers are designed targeting the rfbE sequences (Genbank S83460) of Escherichia coli O157 : H7, and observed the results by electrophoretic and white precipitate. Experiments are carried out on 19 strains of foodborne pathogenic bacteria using LAMP technique, and the results show LAMP-negative expect three E. coli O157 strains. The detection limit is 1.0 CFU/mL for pure culture of E. coli O157, while 1.4 CFU/g is the detection limit for artificially contaminated beef sample. Results demonstrate that LAMP is a simple, sensitive, specific and inexpensive method for the gene detection of E. coli O157. What is more, there is a huge potential for detection of foodborne pathogenic bacteria in the future.