Detection of the van Alphabet and Identification of Enterococci and Staphylococci at the Species Level by Multiplex PCR

Abstract

A multiplex PCR assay was developed for detection of the six types of glycopeptide resistance characterized in enterococci and for identification of Enterococcus faecium , Enterococcus faecalis , Staphylococcus aureus , and Staphylococcus epidermidis at the species level. Primers targeting the genes vanA , vanB , vanC , vanD , vanE , vanG , and ddl of E. faecium and E. faecalis and nuc of S. aureus and a chromosomal portion specific to S. epidermidis were designed to allow amplification of fragments with various sizes. This specific and sensitive technique allows detection of glycopeptide-resistant strains, in particular methicillin-resistant S. aureus , that may escape phenotype-based automated rapid methods.