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.
authors.affiliation: Ibis Biosciences, Inc., 1896 Rutherford Rd., Carlsbad, California 92008, USA.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Hall
authors.forename: Rangarajan
authors.initials: R
authors.lastname: Sampath
authors.forename: Lawrence B
authors.initials: LB
authors.lastname: Blyn
authors.forename: Raymond
authors.initials: R
authors.lastname: Ranken
authors.forename: Cristina
authors.initials: C
authors.lastname: Ivy
authors.forename: Rachael
authors.initials: R
authors.lastname: Melton
authors.forename: Heather
authors.initials: H
authors.lastname: Matthews
authors.forename: Neill
authors.initials: N
authors.lastname: White
authors.forename: Feng
authors.initials: F
authors.lastname: Li
authors.forename: Vanessa
authors.initials: V
authors.lastname: Harpin
authors.forename: David J
authors.initials: DJ
authors.lastname: Ecker
authors.forename: Linda K
authors.initials: LK
authors.lastname: McDougal
authors.forename: Brandi
authors.initials: B
authors.lastname: Limbago
authors.forename: Tracy
authors.initials: T
authors.lastname: Ross
authors.forename: Donna M
authors.initials: DM
authors.lastname: Wolk
authors.forename: Vicki
authors.initials: V
authors.lastname: Wysocki
authors.forename: Karen C
authors.initials: KC
authors.lastname: Carroll
mesh_terms: D015373:Bacterial Typing Techniques / Q000379:methods*; D016000:Cluster Analysis; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013203:Staphylococcal Infections / Q000382:microbiology*; D013211:Staphylococcus aureus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D014481:United States
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial