Development of a single-reaction multiplex PCR toxin typing assay for Staphylococcus aureus strains

We describe here the development of a single-reaction multiplex PCR assay f or the enterotoxin genes from Staphylococcus aureus that utilizes a univers al toxin gene primer in combination,vith toxin-specific primers to amplify characteristic toxin gene products. In combination,with a new DNA purificat ion method, the assay can detect enterotoxin genes A to E from a pure cultu re within 3 to 4 h. The test,vas used to characterize a diverse set of envi ronmental S. aureus isolates, and a 99% correlation with toxin typing using standard immunological tests was found. The design of the assay allows it to be extended to include both newly characterized and as-yet-unknown toxin genes.