ABSTRACT
Objective: To evaluated the PCR for mecA gene compared with the conventional oxacillin disk diffusion method for methicillin-resistant Staphylococcus aureus (S. aureus) identification. Methods: A total of 292 S. aureus strains were isolated from various clinical specimens obtained from hospitalized patients. Susceptibility test to several antimicrobial agents was performed by disk diffusion agar according to Clinical and Laboratory Standards Institute guidelines. The PCR amplification of the mecA gene was carried out in all the clinical isolates.Results:activity and vancomycin was the most effective. The rate of methicillin-resistant S. aureus prevalence determined by oxacillin disk diffusion method was 47.6%; whereas, 45.1% of S. aureus isolates were mecA- positive in the PCR assay. Among antibiotics used in our study, penicillin showed the least anti-staphylococcal Conclusions: This study is suggestive that the PCR for detection of mecA gene is a fast, accurate and valuable diagnostic tool, particularly in hospitals in areas where methicillin-resistant S. aureus is endemic.
ABSTRACT
<p><b>OBJECTIVE</b>To evaluated the PCR for mecA gene compared with the conventional oxacillin disk diffusion method for methicillin-resistant Staphylococcus aureus (S. aureus) identification.</p><p><b>METHODS</b>A total of 292 S. aureus strains were isolated from various clinical specimens obtained from hospitalized patients. Susceptibility test to several antimicrobial agents was performed by disk diffusion agar according to Clinical and Laboratory Standards Institute guidelines. The PCR amplification of the mecA gene was carried out in all the clinical isolates.</p><p><b>RESULTS</b>Among antibiotics used in our study, penicillin showed the least anti-staphylococcal activity and vancomycin was the most effective. The rate of methicillin-resistant S. aureus prevalence determined by oxacillin disk diffusion method was 47.6%; whereas, 45.1% of S. aureus isolates were mecA- positive in the PCR assay.</p><p><b>CONCLUSIONS</b>This study is suggestive that the PCR for detection of mecA gene is a fast, accurate and valuable diagnostic tool, particularly in hospitals in areas where methicillin-resistant S. aureus is endemic.</p>
ABSTRACT
BACKGROUND: Coagulase-negative staphylococci (CNS) has been considered as a major causative agent of nosocomial infections. A prompt and accurate detection of methicillin resistance (MR) in staphylococci is a current issue of clinical microbiology laboratories. This study was purposed to evaluate various methods for detecting MR from CNS. METHODS: We selected 78 CNS strains obtained from blood cultures from April 1999 through July 2001 including 20 strains of Staphylococcus epidermidis, 20 S. hominis (SHO), 19 S. capitis, 9 S. haemolyticus, 3 S. saccharolyticus, 1 S. saprophyticus (SAP), 2 S. warneri (SWA), 2 S. xylosus, 1 S. lugdunensis, and 1 S. auricularis. In addition, one SAP strain received from World Health Organization for proficiency tests was also studied. The following methods were compared to the mecA gene PCR: MicroScan PosCombo 12, oxacillin salt agar containing 6 microgram/mL (OSA-6) or 0.6 microgram/mL (OSA- 0.6) of oxacillin, oxacillin disk diffusion (ODD), and MRSA-Screen latex agglutination (LA) for detecting penicillin binding protein 2a. RESULTS: One SWA was failed in mecA-PCR and fifty-nine of 78 (75.6%) CNS were positive for mecA gene. The agreement rates, sensitivities, and specificities for each test were as follows: for MicroScan, 97.3%, 98.2%, 88.9%; for OSA-6 and OSA-0.6 at 24-h incubation, 79.5%, 74.6%, 94.7% and 79.5%, 72.9%, 100%, respectively, and at 48-h incubation, 91.0%, 91.5%, 89.5% and 91.0%, 96.6%, 73.7%, respectively; ODD, 84.6%, 84.7%, 84.2%; LA, 80.8%, 76.3%, 94.7%. One SHO and one SAP that were mecA-negative showed resistance in the MicroScan, ODD, and OSA. CONCLUSIONS: MicroScan appears a reliable method to detect MR in all species of CNS except SHO and SAP. ODD and LA were not appropriate in detecting MRCNS due to a low sensitivity. Although OSA-0.6 at 48-h incubation showed a high sensitivity, the low specificity may limit a routine use in clinical laboratory.