ORIGINAL ARTICLE

Detection of methicillin and mupirocin resistance in Staphylococcus aureus isolates using conventional and molecular methods: a descriptive study from a burns unit with high prevalence of MRSA

P U Krishnan, K Miles and N Shetty

Department of Clinical Microbiology, University College London Hospitals and PHLS Collaborating Centre, Grafton Way, III Floor, London W1E 6DB, UK

Correspondence to:
Correspondence to:
Dr N Shetty, Department of Clinical Microbiology UCLH-PHLS Collaborating Centre, III Floor Out Patient Building, Grafton Way, London WC1E 6DB, UK; nandini.shetty{at}uclh.org

Aims: To compare conventional phenotypic methods for the detection of methicillin and mupirocin resistance in Staphylococcus aureus in routine laboratory practice with reference to an established molecular method.

Methods: This study was conducted on a selection of 65 isolates of methicillin resistant Staphylococcus aureus (MRSA) from a burns unit in India which is endemic for MRSA. The Kirby–Bauer and modified Stokes disc diffusion tests and the Vitek^TM breakpoint minimum inhibitory concentration (MIC) were performed on all isolates using the presence of the mecA gene as the reference standard. Gel based and colorimetric polymerase chain reaction (PCR) assays were evaluated as molecular methods for the diagnosis of MRSA. A commercial latex agglutination test, the Mastalex^TM, was assessed for the detection of penicillin binding protein 2a (PBP2a), the mecA gene product. Conventional disc diffusion and molecular methods were investigated for the detection of mupirocin resistance.

Results: Fifty one of 65 isolates were positive for the mecA gene. All three phenotypic methods showed high sensitivity (> 96.2%), whereas the specificity varied: 50% for Kirby–Bauer, 87.5% for modified Stokes, and 93.3% for Vitek. The colorimetric PCR was less cumbersome than the gel based PCR; there was complete concordance between both systems. The Mastalex^TM kit showed good correlation with PCR. One isolate was found to be mupirocin resistant and harboured the mupA gene.

Conclusions: The specificity of routine laboratory tests for MRSA detection was variable. mecA gene detection, the "gold standard" to confirm ambiguous results, is difficult to perform in routine diagnostic laboratories. The Mastalex^TM kit for the detection of PBP2a is an alternative that could be used in most laboratories. High level mupirocin resistance can be confirmed with genotypic methods.

Keywords: mecA gene; MRSA; mupirocin; laboratory diagnosis

Abbreviations: BSAC, British Society for Antimicrobial Chemotherapy; ELONA, enzyme linked oligonucleotide assay; HRP, horseradish peroxidase; MIC, minimum inhibitory concentration; MRSA, methicillin resistant Staphylococcus aureus; NCCLS, National Committee for Clinical Laboratory Standards; PBP2a, penicillin binding protein 2a; PCR, polymerase chain reaction; SJMCH, St John’s Medical College Hospital; UCLH, University College London Hospitals; UCLMS, University College London Medical School

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

• Arakere, G., Nadig, S., Swedberg, G., Macaden, R., Amarnath, S. K., Raghunath, D. (2005). Genotyping of Methicillin-Resistant Staphylococcus aureus Strains from Two Hospitals in Bangalore, South India. J. Clin. Microbiol. 43: 3198-3202 [Abstract] [Full Text]