Typing of bovine attaching and effacing Escherichia coli by multiplex in vitro amplification of virulence-associated genes

Attaching and effacing Escherichia coli is a new causal agent of diarrhea in calves. Its major virulence factors are the intimin protein, encoded by the eaeA gene, and the Shiga-like toxins, encoded by sit genes, Because the sequences of these genes are available, we selected specific primers to amplify each virulence gene so as to develop a new identification test based on multiplex amplification of virulence-associated genes. Of 30 tested strains, 14 were eaeA(+), 15 were eneA(+) slt-I+, 1 was eneA(+) slt-I+ slt-II+, and 1 was eaeA(+) slt-II+. The method proved in our hands to be fast and specific and in perfect correlation with the hybridization method.