摘要
应用实时荧光PCR技术建立一种检测冻干奶粉样品中大肠杆菌的方法。通过传统培养法将大肠杆菌O157和O111分离纯化,根据大肠杆菌O157和O111的两对特异性基因rfbE和wbdl设计引物和探针,进行RT-PCR检测;由于该大肠杆菌可能是产志贺毒素大肠杆菌,对STEC的特异性毒力基因stx1、stx2b、eae设计引物和探针并检测。结果表明,该冻干奶粉样品中含有大肠杆菌O157和O111,且致病菌O157检出毒力基因stx1、stx2b、eae,即产志贺毒素与黏附素,致病菌O111检出毒力基因eae,不产志贺毒素而产黏附素。该方法能准确分离大肠杆菌O157和O111,利用实时荧光PCR的高灵敏度和特异性快速检验致病菌及其特异性毒力基因。
The real time fluorescence PCR was applied to establish a detection method for Escherichia coli O157 and O111in lyophilized milk powder.E.coli O157 and O111 were isolated and purified through traditional culture method,and the primers and probes were designed based on the rfbEgene and wbdl gene of E.coli O157 and O111.Considering the possibility of STEC,the primers and probes were designed based on the toxin genes stx1,stx2 b,eae and the genes were detected by RT-PCR.The results showed that lyophilized milk powder was contaminated by E.coli O157 and O111.Stx1 and stx2b genes were detected in E.coli O157 which could produce Shiga toxin and adhesin,while the eae gene was detected in E.coli O111 which could produce adhesin.E.coli O157 and O111could be separated accurately by RT-PCR method,indicating the method could be used in quickly detecting for the pathogens and toxin genes in high sensitivity and specificity.
作者
曹文博
郑秋月
张公亮
侯红漫
CAO Wenbo ZHENG Qiuyue ZHANG Gongliang HOU Hongman(School of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, China Liaoning Entry-Exit Inspection and Quarantine Bureau, Dalian 116001, China)
出处
《大连工业大学学报》
CAS
北大核心
2016年第5期317-320,共4页
Journal of Dalian Polytechnic University
基金
质检公益性行业科研专项(201210043)
