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

doi:10.1128/JCM.02175-08

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

School of Medicine

Research output: Contribution to journal › Article › peer-review

52 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 language	English (US)
Pages (from-to)	1733-1741
Number of pages	9
Journal	Journal of clinical microbiology
Volume	47
Issue number	6
DOIs	https://doi.org/10.1128/JCM.02175-08
State	Published - Jun 2009

ASJC Scopus subject areas

Microbiology (medical)

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10.1128/JCM.02175-08

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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

Hall, TA, Sampath, R, Blyn, LB, Ranken, R, Ivy, C, Melton, R, Matthews, H, White, N, Li, F, Harpin, V, Ecker, DJ, McDougal, LK, Limbago, B, Ross, T, Wolk, DM, Wysocki, V & Carroll, KC 2009, 'Rapid molecular genotyping and clonal complex assignment of Staphylococcus aureus isolates by PCR coupled to electrospray ionization-mass spectrometry', Journal of clinical microbiology, vol. 47, no. 6, pp. 1733-1741. https://doi.org/10.1128/JCM.02175-08

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

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.",

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

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AU - Ivy, Cristina

AU - Melton, Rachael

AU - Matthews, Heather

AU - White, Neill

AU - Li, Feng

AU - Harpin, Vanessa

AU - Ecker, David J.

AU - McDougal, Linda K.

AU - Limbago, Brandi

AU - Ross, Tracy

AU - Wolk, Donna M.

AU - Wysocki, Vicki

AU - Carroll, Karen C.

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

Abstract

ASJC Scopus subject areas

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