Detection of Shiga-Like Toxin (stx(1) and stx(2)), Intimin (eaeA), and Enterohemorrhagic Escherichia coli (EHEC) Hemolysin (EHEC hlyA) Genes in Animal Feces by Multiplex PCR

A multiplex PCR was developed for the rapid detection of genes encoding Shiga toxins 1 and 2 (stx(1) and stx(2)), intimin (eaeA), and enterohemolysin A (hlyA) in 444 fecal samples derived from healthy and clinically affected cattle, sheep, pigs, and goats. The method involved non-solvent-based extraction of nucleic acid from an aliquot of an overnight culture of feces in EC (modified) broth. The detection limit of the assay for both fecal samples and pure cultures was between 18 and 37 genome equivalents. stx(1) and hlyA were the most commonly encountered virulence factors.