The combination of CRISPR-MVLST and PFGE provides increased discriminatory power for differentiating human clinical isolates of Salmonella enterica subsp enterica serovar Enteritidis
The combination of CRISPR-MVLST and PFGE provides increased discriminatory power for differentiating human clinical isolates of Salmonella enterica subsp enterica serovar Enteritidis
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DOI：
10.1016/j.fm.2012.11.012
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发表时间：
2013-05-01
影响因子：
5.3
通讯作者：
Dudley, Edward. G.
中科院分区：
文献类型：
--
作者：
Shariat, Nikki;DiMarzio, Michael J.;Dudley, Edward. G.
肠道沙门氏菌肠道亚种enterica serovar Enterilis（S.肠杆菌病）是食源性沙门氏菌病的主要原因。为了追踪特定的沙门氏菌菌株，需要快速、高效、准确的鉴定方法。人类沙门氏菌病暴发期间的肠道感染。通过利用毒力基因的高变性质和重复的规则间隔短回文重复序列（CRISPR），我们先前开发了一种强大的基于序列的亚型分型方法，称为CRISPR-MVLST。为了证实CRISPR-MVLST的适用性，我们分析了一组广泛的S.在六年期间收集的肠杆菌分离株。在141个分离株中，我们定义了22个肠毒素序列类型（EST），其中大多数是新的。值得注意的是，菌株表现出共同的PFGE模式，JEGX 01.0004（特征为与40%的S.美国的肠杆菌分离株）被分成12种不同的序列类型。相反，EST 4的分离株，我们观察到的最主要的EST，包括八个不同的PFGE模式。重要的是，我们发现，一些以前与农场层面食品供应链相关的基因型现在已经在临床样本中得到了鉴定。CRISPR序列数据显示，S的不同等位基因之间存在微妙但明显的差异。肠杆菌，这表明这些位点的进化是垂直发生的，而不是以前报道的进化间隔收购其他细菌。(C)2012爱思唯尔有限公司保留所有权利。
Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) is a major cause of foodborne salmonellosis. Rapid, efficient and accurate methods for identification are required to track specific strains of S. Enteritidis during outbreaks of human salmonellosis. By exploiting the hypervariable nature of virulence genes and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs), we previously developed a powerful sequence-based subtyping approach, designated CRISPR-MVLST. To substantiate the applicability of CRISPR-MVLST, we analyzed a broad set of S. Enteritidis isolates collected over a six-year period. Among 141 isolates we defined 22 Enteritidis Sequence Types (ESTs), the majority of which were novel. Notably, strains exhibiting the common PFGE pattern, JEGX01.0004 (characteristic of similar to 40% of S. Enteritidis isolates in the United States), were separated into twelve distinct sequence types. Conversely, isolates of EST4, the most predominant EST we observed, comprised eight different PFGE patterns. Importantly, we showed that some genotypes that were previously associated with the food supply chain at the farm level have now been identified in clinical samples.CRISPR sequence data shows subtle but distinct differences among different alleles of S. Enteritidis, suggesting that evolution of these loci occurs vertically, as opposed to previously reported evolution by spacer acquisition in other bacteria. (C) 2012 Elsevier Ltd. All rights reserved.