Teagasc, Ashtown Food Research Centre, Ashtown, Dublin 15, Ireland.
J Appl Microbiol. 2009 Oct;107(4):1340-9. doi: 10.1111/j.1365-2672.2009.04320.x. Epub 2009 Apr 18.
To determine the degree of relatedness between isolates of Escherichia coli O157:H7 of human, bovine, ovine and porcine origin.
Escherichia coli O157:H7 isolates were compared using (i) PFGE XbaI patterns, (ii) PCR profiles of virulence genes and (iii) the DNA sequences of genes reported to play a role in pathogenicity. The 77 E. coli O157:H7 isolates demonstrated 49 different PFGE patterns of which, eight were common to multiple isolates, and the remaining 41 were distinct. Isolates of different origin did not correlate, except for one cluster consisting of two human and two beef isolates. The majority of animal isolates had the same PCR profiles of virulence genes as those isolated from clinical patients. Single nucleotide polymorphisms (SNPs) were identified in the sequence of a 255-bp region of the vtx2 subunit A gene.
Six SNPs were detected in the vtx2A gene, defining four different haplotypes. One nonsynonymous substitution encoded for an amino acid change from glutamic to aspartic acid.
Results indicate that although E. coli O157:H7 isolates of differing origin were distinct by PFGE, the DNA sequences of the main virulence genes associated with human clinical illness were conserved.
确定人源、牛源、羊源和猪源大肠杆菌 O157:H7 分离株之间的亲缘关系程度。
使用(i)XbaI 脉冲场凝胶电泳(PFGE)模式、(ii)毒力基因 PCR 图谱和(iii)据报道与致病性相关的基因 DNA 序列,对大肠杆菌 O157:H7 分离株进行了比较。77 株大肠杆菌 O157:H7 分离株显示出 49 种不同的 XbaI PFGE 模式,其中 8 种模式在多个分离株中共同存在,其余 41 种模式则各不相同。不同来源的分离株之间没有相关性,除了由两个人类和两个牛肉分离株组成的一个聚类外。大多数动物分离株的毒力基因 PCR 图谱与从临床患者中分离出的毒力基因相同。在 vtx2 亚单位 A 基因的 255bp 区域的序列中发现了单核苷酸多态性(SNP)。
在 vtx2A 基因中检测到 6 个 SNP,定义了 4 种不同的单倍型。一个非同义突变导致谷氨酸到天冬氨酸的氨基酸变化。
结果表明,尽管来自不同来源的大肠杆菌 O157:H7 分离株在 PFGE 上是不同的,但与人类临床疾病相关的主要毒力基因的 DNA 序列是保守的。