Multiplex PCR for simultaneous identification of Staphylococcus aureus and detection of methicillin and mupirocin resistance.
In this work, we describe a multiplex PCR assay for the detection of clinically relevant antibiotic resistance genes harbored by some Staphylococcus aureus isolates and for the simultaneous identification of such isolates at the species level. Conditions were optimized for the simultaneous detection of the 310-, 456-, and 651-bp regions of the mecA (encoding high-level methicillin resistance), ileS-2 (encoding high-level mupirocin resistance), and femB (encoding a factor essential for methicillin resistance) genes, respectively, from a single colony in a single reaction tube. The femB PCR fragment allows the specific identification of S. aureus. Validation of the method was performed using 50 human isolates of methicillin-resistant S. aureus (MRSA) and the appropriate control strains. This assay offers a rapid, simple, feasible, specific, sensitive, and accurate identification of mupirocin-resistant MRSA clinical isolates and could be systematically applied as a diagnostic test in clinical microbiology laboratories, facilitating the design and use of antibiotic therapy.
Pérez-Roth
E
E
Molecular Biology Laboratory, Research Unit, Nuestra Señora de Candelaria Hospital, Santa Cruz de Tenerife, Spain.
Claverie-Martín
F
F
Villar
J
J
Méndez-Alvarez
S
S
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D006801:Humans; D016106:Methicillin Resistance / Q000235:genetics; D008826:Microbial Sensitivity Tests; D016712:Mupirocin / Q000494:pharmacology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013203:Staphylococcal Infections / Q000382:microbiology; D013211:Staphylococcus aureus / Q000145:classification* / Q000187:drug effects* / Q000235:genetics
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004269:DNA, Bacterial; C102928:FemB protein, Staphylococcus epidermidis; D016712:Mupirocin