Multiplex PCR assay to identify methicillin-resistant Staphylococcus haemolyticus.
Staphylococcus haemolyticus is the most frequently coagulase-negative Staphylococcus species associated with antimicrobial resistance isolated from nosocomial infections. We developed an accurate and simple multiplex PCR assay to identify methicillin-resistant S. haemolyticus (MRSH) isolates. We designed species-specific primers of the mvaA gene that encodes a 3-hydroxy-3-methylglutaryl coenzyme A involved in the mevalonate pathway of the microorganism. Simultaneously, mecA gene primers of methicillin resistance were also used. The PCR assay was established using 16 strains of different reference Staphylococcus species and validated with a collection of 147 clinical staphylococcal isolates that were also phenotypically characterized. Reliable results for the detection of MRSH isolates were obtained for 100% of the strains evaluated, showing that this PCR assay can be used for the routine microbiology laboratories. This is the first report using species-specific multiplex PCR to detect a single segment of S. haemolyticus associated with a segment of mecA gene.
Schuenck
Ricardo P
RP
Department of Medical Microbiology, Institute of Microbiology, Rio de Janeiro Federal University, Rio de Janeiro, Brazil.
Pereira
Eliezer M
EM
Iorio
Natalia L P
NL
Dos Santos
Kátia R N
KR
D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D008712:Methicillin / Q000494:pharmacology; D016106:Methicillin Resistance; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013203:Staphylococcal Infections / Q000175:diagnosis / Q000382:microbiology; D041162:Staphylococcus haemolyticus / Q000145:classification* / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification
D017931:DNA Primers; D004269:DNA, Bacterial; D008712:Methicillin