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Journal of Clinical Microbiology, May 2008, p. 1620-1627, Vol. 46, No. 5
0095-1137/08/$08.00+0     doi:10.1128/JCM.02453-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Validation of Virulence and Epidemiology DNA Microarray for Identification and Characterization of Staphylococcus aureus Isolates{triangledown} ,{dagger}

Richard P. Spence, Victoria Wright, Dlawer A. A. Ala-Aldeen, David P. Turner, Karl G. Wooldridge, and Richard James*

Centre for Healthcare Associated Infections, Institute of Infection, Immunity and Inflammation, CBS Building, University Park, University of Nottingham, Nottingham NG7 2RD, United Kingdom

Received 20 December 2007/ Returned for modification 29 December 2007/ Accepted 13 February 2008

The human pathogen Staphylococcus aureus is isolated and characterized using traditional culture and sensitivity methodologies that are slow and offer limited information on the organism. In contrast, DNA microarray technology can provide detailed, clinically relevant information on the isolate by detecting the presence or absence of a large number of virulence-associated genes simultaneously in a single assay. We have developed and validated a novel, cost-effective multiwell microarray for the identification and characterization of Staphylococcus aureus. The array comprises 84 gene targets, including species-specific, antibiotic resistance, toxin, and other virulence-associated genes, and is capable of examining 13 different isolates simultaneously, together with a reference control strain. Analysis of S. aureus isolates whose complete genome sequences have been determined (Mu50, N315, MW2, MRSA252, MSSA476) demonstrated that the array can reliably detect the combination of genes known to be present in these isolates. Characterization of a further 43 S. aureus isolates by the microarray and pulsed-field gel electrophoresis has demonstrated the ability of the array to differentiate between isolates representative of a spectrum of S. aureus types, including methicillin-susceptible, methicillin-resistant, community-acquired, and vancomycin-resistant S. aureus, and to simultaneously detect clinically relevant virulence determinants.


* Corresponding author. Mailing address: Centre for Healthcare Associated Infections, Institute of Infection, Immunity and Inflammation, CBS Building, University Park, University of Nottingham, Nottingham NG7 2RD, United Kingdom. Phone: 44 115 84667952. Fax: 44 115 8467951. E-mail: richard.james{at}nottingham.ac.uk

{triangledown} Published ahead of print on 20 February 2008.

{dagger} Supplemental material for this article may be found at http://jcm.asm.org/.


Journal of Clinical Microbiology, May 2008, p. 1620-1627, Vol. 46, No. 5
0095-1137/08/$08.00+0     doi:10.1128/JCM.02453-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.







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