Rapid molecular genotyping and clonal complex assignment of Staphylococcus aureus isolates by PCR coupled to electrospray ionization-mass spectrometry

Thomas A. Hall, Rangarajan Sampath, Lawrence B. Blyn, Raymond Ranken, Cristina Ivy, Rachael Melton, Heather Matthews, Neill White, Feng Li, Vanessa Harpin, David J. Ecker, Linda K. McDougal, Brandi Limbago, Tracy Ross, Donna M. Wolk, Vicki Wysocki, Karen C. Carroll

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50 Scopus citations

Abstract

We describe a high-throughput assay using PCR coupled to electrospray ionization-mass spectrometry (PCR/ESI-MS) to determine the genotypes of Staphylococcus aureus isolates. The primer sets used in the PCR/ESI-MS assay were designed to amplify the same genes analyzed in multilocus sequence typing (MLST). The method was used to identify the clonal complex and USA type of each isolate and is suitable for use in a clinical or public-health setting. The method was validated using a panel of diverse isolates from the Centers for Disease Control and Prevention that were previously characterized by MLST and pulsed-field gel electrophoresis (PFGE). Clinical isolates from two geographically distinct hospitals were characterized, and the clustering results were in agreement with those for repetitive-element PCR and PFGE. The PCR/ESI-MS method enables genotyping of over 180 samples of S. aureus per day in an automated fashion.

Original languageEnglish (US)
Pages (from-to)1733-1741
Number of pages9
JournalJournal of clinical microbiology
Volume47
Issue number6
DOIs
StatePublished - Jun 1 2009

ASJC Scopus subject areas

  • Microbiology (medical)

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    Hall, T. A., Sampath, R., Blyn, L. B., Ranken, R., Ivy, C., Melton, R., Matthews, H., White, N., Li, F., Harpin, V., Ecker, D. J., McDougal, L. K., Limbago, B., Ross, T., Wolk, D. M., Wysocki, V., & Carroll, K. C. (2009). Rapid molecular genotyping and clonal complex assignment of Staphylococcus aureus isolates by PCR coupled to electrospray ionization-mass spectrometry. Journal of clinical microbiology, 47(6), 1733-1741. https://doi.org/10.1128/JCM.02175-08