Ison Sarah A, Delannoy Sabine, Bugarel Marie, Nightingale Kendra K, Webb Hattie E, Renter David G, Nagaraja Tiruvoor G, Loneragan Guy H, Fach Patrick
Texas Tech University, Department of Animal and Food Sciences, Lubbock, Texas, USA.
French Agency for Food, Environmental and Occupational Health and Safety, Food Safety Laboratory, Platform IdentyPath, Maisons-Alfort, France.
Appl Environ Microbiol. 2015 Jun;81(11):3671-8. doi: 10.1128/AEM.00397-15. Epub 2015 Mar 20.
Escherichia coli O26 has been identified as the most common non-O157 Shiga toxin-producing E. coli (STEC) serogroup to cause human illnesses in the United States and has been implicated in outbreaks around the world. E. coli has high genomic plasticity, which facilitates the loss or acquisition of virulence genes. Attaching and effacing E. coli (AEEC) O26 strains have frequently been isolated from bovine feces, and there is a need to better characterize the relatedness of these strains to defined molecular pathotypes and to describe the extent of their genetic diversity. High-throughput real-time PCR was used to screen 178 E. coli O26 isolates from a single U.S. cattle feedlot, collected from May to July 2011, for the presence or absence of 25 O26 serogroup-specific and virulence-associated markers. The selected markers were capable of distinguishing these strains into molecularly defined groups (yielding 18 unique marker combinations). Analysis of the clustered regularly interspaced short palindromic repeat 1 (CRISPR1) and CRISPR2a loci further discriminated isolates into 24 CRISPR types. The combination of molecular markers and CRISPR typing provided 20.8% diversity. The recent CRISPR PCR target SP_O26-E, which was previously identified only in stx2-positive O26:H11 human clinical strains, was identified in 96.4% (161/167 [95% confidence interval, 99.2 to 93.6%]) of the stx-negative AEEC O26:H11 bovine fecal strains. This supports that these stx-negative strains may have previously contained a prophage carrying stx or could acquire this prophage, thus possibly giving them the potential to become pathogenic to humans. These results show that investigation of specific genetic markers may further elucidate our understanding of the genetic diversity of AEEC O26 strains in bovine feces.
大肠杆菌O26已被确定为美国导致人类疾病的最常见非O157产志贺毒素大肠杆菌(STEC)血清群,并在世界各地的疫情中被牵连。大肠杆菌具有高度的基因组可塑性,这有利于毒力基因的丢失或获得。黏附性大肠杆菌(AEEC)O26菌株经常从牛粪中分离出来,需要更好地描述这些菌株与已定义分子致病型的相关性,并描述其遗传多样性的程度。使用高通量实时PCR对2011年5月至7月从美国一个单一的肉牛饲养场收集的178株大肠杆菌O26分离株进行筛选,以检测25种O26血清群特异性和毒力相关标记的存在与否。所选标记能够将这些菌株分为分子定义的组(产生18种独特的标记组合)。对成簇规律间隔短回文重复序列1(CRISPR1)和CRISPR2a位点的分析进一步将分离株分为24种CRISPR类型。分子标记和CRISPR分型的组合提供了20.8%的多样性。最近的CRISPR PCR靶点SP_O26-E,以前仅在stx2阳性的O26:H11人类临床菌株中发现,在96.4%(161/167[95%置信区间,99.2至93.6%])的stx阴性AEEC O26:H11牛粪菌株中被发现。这支持这些stx阴性菌株可能以前含有携带stx的原噬菌体或可以获得这种原噬菌体,因此可能使它们有对人类致病的潜力。这些结果表明,对特定遗传标记的研究可能会进一步阐明我们对牛粪中AEEC O26菌株遗传多样性的理解。