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当前位置: 首页 > 文章 > 牛源大肠杆菌O157:H7的分离及毒力基因鉴定 中国兽医学报 2012,32 (8) 1148-1153
Position: Home > Articles > Isolation of Escherichia coli O157:H7 from cattle and detection of its virulence genes Chinese Journal of Veterinary Science 2012,32 (8) 1148-1153

牛源大肠杆菌O157:H7的分离及毒力基因鉴定

作  者：
丁浩;苏战强;夏利宁;王雨朦;王栋;张瑾瑜;张桃;况玲;姚刚
单  位：
新疆农业大学动物医学学院
关键词：
牛;肠出血性大肠杆菌O157:H7;毒力基因;致病性
摘  要：
从2个牛场采集新鲜粪便,增菌后,免疫磁珠富集,涂布筛选性培养基,挑取可疑菌落用rfbE/fliC二重PCR和血清学方法鉴定。设计毒力基因stx1、stx2、eae、hlyA和tccp相应引物,针对O157:H7对分离株进行PCR鉴定。口服攻毒链霉素处理的BALB/c小鼠明确分离株致病性。结果显示,成功分离到7株出血性大肠杆菌O157:H7,并且有1株迟缓性发酵山梨醇麦康凯培养基。毒力基因检测显示,其中6株毒力因子表型为stx1-stx2+eae+hlyA+tccp+,另有1株表现型为stx1+stx2+eae+hlyA+tccp+,各分离株tccp基因均为阳性,但携带的重复片段数量有差异。所采集样品中肠出血性大肠杆菌O157:H7的检出率高达12%。1×1010 CFU同剂量口服接种经PBS洗涤的5株O157:H7分离株全菌,小鼠存活率有差异分别为40%,50%,60%,20%,50%,各分离株在小鼠体内排菌时间也有差异分别为攻毒后7,9,13,13,15d。
译  名：
Isolation of Escherichia coli O157:H7 from cattle and detection of its virulence genes
作  者：
YE Qing1,2,ZHANG Xue-han1,HE Kong-wang1,FEI Rong-mei2,ZHAO Pan-deng1,LUAN Xiao-ting1(1.National Center for Engineering Research of Veterinary Bio-products,Key Laboratory of Engineering Research of Veterinary Bio-products of Agricultural Ministry,Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;2.College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China)
关键词：
cattle;enterohaemorrhagic Escherichia coli O157:H7;virulence genes;pathogenicity
摘  要：
To understand the epidemic status of Escherichia coli O157:H7,we collected the feces and isolated O157: H7 strains from cattle farms and detected several major virulent genes.Fresh fecal samples were collected,cultured at 37℃,enriched with immunomagnetic beads and spread onto CHROM agar.After culture at 37℃ for 12 h,suspected colonies were picked and identified with rfbE/fliC PCR and serological methods.The stx1,stx2,eae,hlyA,and tccp primers were designed to amplify the fragments.Oral attack streptomycin treatment of BALB/c mice were determined pathogenic of strains.The results showed that seven O157:H7 isolates were successfully isolated,and No.2 O157:H7 gradually fermented sorbitol.After all isolates were detected major virulence genes,No.6 phenotypic virulence factors were stx1-stx2+eae+hlyA+tccp+,the remaining six isolates were stx1+stx2+eae+hlyA+tccp+.Although all the isolates have all the detected genes,they have different tccp gene that contains different numbers of repeat fragments.This study showed that the collected cattle feces carried O157:H7 and the detection rate is up to 12%.In the mice inoculated with 1×1010CFU dose of PBS washing of five strains of O157: H7 strains isolated their survival rates were 40%,50%,60%,20%,50%,and discharge time in mice were 7,9,13,13,15 days,respectively.

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