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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Authors: | Nikki Shariat Michael J. DiMarzio Shuang Yin Lisa Dettinger Carol H. Sandt James R. Lute Rodolphe Barrangou Edward G. Dudley |
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Affiliation: | 1. Department of Food Science, 326 Food Science Building, The Pennsylvania State University, University Park, PA 16802, USA;2. Division of Clinical Microbiology, The Pennsylvania Department of Health, 110 Pickering Way, Exton, PA 19341, USA;3. DuPont Nutrition and Health, 3329 Agriculture Drive, Madison, WI 53716, USA |
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Abstract: | 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 ∼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. |
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Keywords: | Salmonella Enteritidis CRISPR-MVLST Molecular subtyping |
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