Multiplex PCR for simultaneous detection of enterococcal genes vanA and vanB and staphylococcal genes mecA, ileS-2 and femB.
The experimental transfer of the vanA gene cluster from Enterococcus faecalis to Staphylococcus aureus has raised fears about the occurrence of such genetic transfer in clinical isolates of methicillin-resistant staphylococci. Recently, infections by a S. aureus strain carrying the enterococcal vancomycin resistance vanA gene cluster were reported. The possible emergence and dissemination of these strains is a serious health threat and makes optimization of prevention strategies and fast detection methods absolutely necessary. In the present study, we developed a PCR protocol for simultaneous detection of enterococcal vanA and vanB genes, the staphylococcal methicillin and mupirocin resistance markers mecA and ileS-2, and identification of S. aureus. As no vancomycin-resistant S. aureus isolates were available for our study, we used mixtures of enterococcal and staphylococcal colonies that harbored the different resistance markers to show that these genes could be detected simultaneously. This protocol could be used to facilitate the detection and identification of predictable S. aureus or methicillin-resistant strains carrying vanA or vanB.
Ramos-Trujillo
Elena
E
Hospital Universitario Nuestra Señora de Candelaria, Unidad de Investigación, Carretera del Rosario s/n, 38010 Santa Cruz de Tenerife, Spain.
Pérez-Roth
Eduardo
E
Méndez-Alvarez
Sebastián
S
Claverie-Martín
Félix
F
D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D019729:Carbon-Oxygen Ligases / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D013293:Enterococcus faecalis / Q000235:genetics*; D022761:Gene Transfer, Horizontal; D005798:Genes, Bacterial; D016106:Methicillin Resistance / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D013211:Staphylococcus aureus / Q000235:genetics*; D020713:Vancomycin Resistance / Q000235:genetics
D001426:Bacterial Proteins; D017931:DNA Primers; C102928:FemB protein, Staphylococcus epidermidis; C071437:VanA ligase, Bacteria; C079913:VanB protein, Enterococcus; D019729:Carbon-Oxygen Ligases