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Micro-array for the identification of Shiga toxin-producing Escherichia coli (STEC) seropathotypes associated with Hemorrhagic Colitis and Hemolytic Uremic Syndrome in humans

Bugarel, Marie, Beutin, Lothar, Martin, Annett, Gill, Alexander, Fach, Patrick
International journal of food microbiology 2010 v.142 no.3 pp. 318-329
enterohemorrhagic Escherichia coli, bacterial contamination, microarray technology, food analysis, food contamination, serotypes, genes, Shiga-like toxins, Shiga toxin, virulence, human diseases, hemolytic uremic syndrome, colitis
A micro-array has been developed, based on the GeneDisc® array, for the genetic identification of 12 O-types and 7 H-types of Shiga toxin-producing Escherichia coli (STEC) including the most clinically relevant enterohemorrhagic E. coli (EHEC) serotypes. The genes selected for determination of the O antigens (rfbE O₁₅₇, wzx O₂₆, wzx O₁₀₃, wbd1 O₁₁₁, ihp1 O₁₄₅, wzx O₁₂₁, wzy O₁₁₃, wzy O₉₁, wzx O₁₀₄, wzy O₁₁₈, wzx O₄₅, and wbgN O₅₅) and H-types (fliC H₂, fliC H₇, fliC H₈, fliC H₁₁, fliC H₁₉, fliC H₂₁, and fliC H₂₈) showed a high specificity and concordance with serology. The micro-array also had a high specificity for EHEC-associated virulence factors, including Shiga toxins 1 and 2 (stx1 and stx2), intimin (eae), enterohemolysin (ehxA), serine protease (espP), catalase peroxidase (katP), the type II secretion system (etpD), subtilase cytotoxin (subA), autoagglutinating adhesin (Saa) and type III secreted effectors encoded in the genomic islands OI-122 (ent/espL2, nleB, and nleE) and OI-71 (nleF, nleH1-2, and nleA). The eae gene was detected in all typical EHEC strains, and the pattern of nle genes encoded in OI-71 and OI-122 was found to be closely associated with certain serotypes of typical EHEC and emerging EHEC strains. Virulence plasmid associated genes such as katP, espP, and etpD were more common in EHEC than in STEC strains; this supports their association with virulence. This array constitutes a valuable approach for the identification of STEC strains with a high potential for human virulence.