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在美国国家奶牛小母牛评估项目(1992 - 1992年)期间,对从奶牛犊中分离出的大肠杆菌O157:H7进行脉冲场凝胶电泳基因组分析。

Genomic analysis using pulsed-field gel electrophoresis of Escherichia coli O157: H7 isolated from dairy calves during the United State National Dairy Heifer Evaluation Project (1992-1992).

作者信息

Lee M S, Kaspar C W, Brosch R, Shere J, Luchansky J B

机构信息

Department of Food Microbiology and Toxicology University of Wisconsin, Madison 53706-1187, USA.

出版信息

Vet Microbiol. 1996 Feb;48(3-4):223-30. doi: 10.1016/0378-1135(95)00135-2.

Abstract

The genomic fingerprints of 26 Escherichia coli O157:H7 isolates from calves on 20 farms in 16 states were determined by pulsed-field gel electrophoresis (PFGE). Digestion of genomic DNAs with the restriction enzymes SfiI and XbaI yielded 14 and 18 restriction endonuclease digestion profiles (REDP), respectively. Seventeen farms (85%) had E. coli O157:H7 with a unique REDP, and when more than one calf tested positive on a farm, the isolates displayed identical REDP. Isolates from different farms within the same state displayed distinct REDP, as did most isolates from farms in different states. The exceptions were three farms in New York, Ohio, and Washington that had calves harboring E. coli O157:H7 with the same REDP. In addition to REDP, the toxin profiles of all 26 isolates were determined using oligonucleotide probes to Shiga-like toxins (SLT) I and II. Nineteen (73%) of the E. coli O157:H7 isolates harbored the genes for both SLT and II, while the remaining seven isolates (27%) had the gene for SLT II only. Also, isolates with the same REDP had the same toxin profile. The genomic relatedness among the E. coli O157:H7 isolates was also determined by principal component analysis of Dice similarity indices of REDP. Three clusters were identified, but none of these were associated with a geographic region or toxin profile.

摘要

采用脉冲场凝胶电泳(PFGE)技术对来自16个州20个农场的26株犊牛源大肠杆菌O157:H7分离株进行了基因组指纹图谱分析。用限制性内切酶SfiI和XbaI消化基因组DNA,分别产生了14种和18种限制性内切酶消化图谱(REDP)。17个农场(85%)的大肠杆菌O157:H7具有独特的REDP,当一个农场有多头犊牛检测呈阳性时,分离株显示出相同的REDP。同一州内不同农场的分离株显示出不同的REDP,不同州农场的大多数分离株也是如此。例外情况是纽约、俄亥俄和华盛顿的三个农场,其犊牛携带的大肠杆菌O157:H7具有相同的REDP。除了REDP,还使用针对志贺样毒素(SLT)I和II的寡核苷酸探针测定了所有26株分离株的毒素谱。19株(73%)大肠杆菌O157:H7分离株同时携带SLT I和II基因,其余7株(27%)仅携带SLT II基因。此外,具有相同REDP的分离株具有相同的毒素谱。还通过对REDP的Dice相似性指数进行主成分分析,确定了大肠杆菌O157:H7分离株之间的基因组相关性。鉴定出三个聚类,但这些聚类均与地理区域或毒素谱无关。

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