A comparison of PCR detection of mecA with oxacillin disk susceptibility testing in different media and sceptor automated system for both Staphylococcus aureus and coagulase-negative staphylococci isolates.
OBJECTIVE
To evaluate three methods for 406 isolates of Staphylococcus aureus and coagulase-negative staphylococci (CNS) for the detection of methicillin resistance (MR) using National Committee for Clinical Laboratory Standards (NCCLS) new interpretive criteria.


METHODS
We used polymerase chain reaction (PCR) as a gold standard method to evaluate three methods [disk diffusion with Mueller-Hinton agar (MHA) and mannitol salt agar (MSA) and Sceptor system (Becton Dickinson, USA)] for the detection of mecA gene. The isolates that were methicillin-resistant with any of the three tests were evaluated further for MR by E-test.


RESULTS
MHA, MSA and Sceptor showed sensitivities of 100, 100 and 99% for S. aureus and 100, 82.6 and 72.1% for CNS, respectively. The specificities of the same methods were found as 100, 90.1 and 99.3% for S. aureus and 79.2, 95.8 and 97.2% for CNS, respectively. E-test showed 100% sensitivity for both S. aureus and CNS. Forty-eight CNS and 16 S. aureus isolates, which presented discrepancies with the three phenotypic methods (MHA disk diffusion method, MSA disk diffusion method and Sceptor), were correctly classified as resistant/susceptible with the E-test when compared with PCR. Only five CNS isolates, which were mecA-negative with PCR were resistant with E-test. Analysis of 248 S. aureus revealed that MHA is superior to other phenotype-based susceptibility testing methods in detecting MR. When we examined the results of 158 CNS, none of the three methods proved efficient in detecting MR.


CONCLUSIONS
We conclude that although the accuracy of the MHA disk diffusion test for the detection of MR approaches the accuracy of PCR for S. aureus isolates, the need for easy and reliable methods of detecting MR in CNS still remains.
Ercis
S
S
Department of Microbiology and Clinical Microbiology, Hacettepe University Faculty of Medicine, Ankara, Turkey. sercis@hacettepe.edu.tr
Sancak
B
B
Hasçelik
G
G
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics*; D003470:Culture Media; D004269:DNA, Bacterial / Q000235:genetics; D006801:Humans; D016106:Methicillin Resistance / Q000235:genetics* / Q000502:physiology; D008826:Microbial Sensitivity Tests / Q000379:methods*; D010068:Oxacillin / Q000494:pharmacology*; D046915:Penicillin-Binding Proteins; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013203:Staphylococcal Infections / Q000382:microbiology; D013210:Staphylococcus / Q000187:drug effects* / Q000235:genetics* / Q000302:isolation & purification
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D003470:Culture Media; D004269:DNA, Bacterial; C487495:MecA protein, Staphylococcus epidermidis; D046915:Penicillin-Binding Proteins; C487789:mecA protein, Staphylococcus aureus; D010068:Oxacillin