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Journal of Clinical Microbiology, October 2009, p. 3129-3137, Vol. 47, No. 10
0095-1137/09/$08.00+0     doi:10.1128/JCM.00709-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Pathogen Profiling: Rapid Molecular Characterization of Staphylococcus aureus by PCR/Electrospray Ionization-Mass Spectrometry and Correlation with Phenotype{triangledown}

Donna M. Wolk,1* Lawrence B. Blyn,2 Thomas A. Hall,2 Rangarajan Sampath,2 Raymond Ranken,2 Cristina Ivy,2 Rachael Melton,2 Heather Matthews,2 Neill White,2 Feng Li,2 Vanessa Harpin,2 David J. Ecker,2 Brandi Limbago,3 Linda K. McDougal,3 Vicki H. Wysocki,1 Mian Cai,4 and Karen C. Carroll4

University of Arizona/BIO5 Institute, 1501 N. Campbell Ave., Tucson, Arizona,1 Ibis Biosciences, Abbott Molecular Inc., 1896 Rutherford Rd., Carlsbad, California 92008,2 Division of Healthcare Quality Promotion, Centers for Disease Control and Prevention, Atlanta, Georgia 30333,3 Johns Hopkins University, 600 N. Wolfe Street, Baltimore, Maryland 212874

Received 7 April 2009/ Returned for modification 2 June 2009/ Accepted 17 August 2009

There are few diagnostic methods that readily distinguish among community-acquired methicillin (meticillin)-resistant Staphylococcus aureus strains, now frequently transmitted within hospitals. We describe a rapid and high-throughput method for bacterial profiling of staphylococcal isolates. The method couples PCR to electrospray ionization-mass spectrometry (ESI-MS) and is performed on a platform suitable for use in a diagnostic laboratory. This profiling technology produces a high-resolution genetic signature indicative of the presence of specific genetic elements that represent distinctive phenotypic features. The PCR/ESI-MS signature accurately identified genotypic determinants consistent with phenotypic traits in well-characterized reference and clinical isolates of S. aureus. Molecular identification of the antibiotic resistance genes correlated strongly with phenotypic in vitro resistance. The identification of toxin genes correlated with independent PCR analyses for the toxin genes. Finally, isolates were correctly classified into genotypic groups that correlated with genetic clonal complexes, repetitive-element-based PCR patterns, or pulsed-field gel electrophoresis types. The high-throughput PCR/ESI-MS assay should improve clinical management of staphylococcal infections.

* Corresponding author. Mailing address: Department of Pathology, University of Arizona, 1501 N. Campbell Ave., P.O. Box 245059, Tucson, AZ 85724-5059. Phone: (520) 626-3676. Fax: (520) 694-7329. E-mail: dwolk{at}email.arizona.edu

{triangledown} Published ahead of print on 26 August 2009.

Journal of Clinical Microbiology, October 2009, p. 3129-3137, Vol. 47, No. 10
0095-1137/09/$08.00+0     doi:10.1128/JCM.00709-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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