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Current methods for screening Enterohemorrhagic  Escherichia coli  (EHEC) O157 and non-O157 in beef enrichments typically rely on the molecular detection of  stx, eae , and serogroup-specific  wzx  or  wzy  gene fragments. As these genetic markers can also be found in some non-EHEC strains, a number of “false positive” results are obtained. Here, we explore the suitability of five novel molecular markers,  espK, espV, ureD, Z2098 , and CRISPR O26:H11  as candidates for a more accurate screening of EHEC strains of greater clinical significance in industrialized countries. Of the 1739 beef enrichments tested, 180 were positive for both  stx  and  eae  genes. Ninety (50%) of these tested negative for  espK, espV, ureD , and  Z2098 , but 12 out of these negative samples were positive for the CRISPR O26:H11  gene marker specific for a newly emerging virulent EHEC O26:H11 French clone. We show that screening for  stx, eae, espK , and  espV , in association with the CRISPR O26:H11  marker is a better approach to narrow down the EHEC screening step in beef enrichments. The number of potentially positive samples was reduced by 48.88% by means of this alternative strategy compared to the European and American reference methods, thus substantially improving the discriminatory power of EHEC screening systems. This approach is in line with the EFSA (European Food Safety Authority) opinion on pathogenic STEC published in 2013.

Revisiting the STEC Testing Approach: Using espK and espV to Make Enterohemorrhagic Escherichia coli (EHEC) Detection More Reliable in Beef