Rapid molecular genotyping and clonal complex assignment of Staphylococcus aureus isolates by PCR coupled to electrospray ionization-mass spectrometry.
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.
Hall
Thomas A
TA
Ibis Biosciences, Inc., 1896 Rutherford Rd., Carlsbad, California 92008, USA.
Sampath
Rangarajan
R
Blyn
Lawrence B
LB
Ranken
Raymond
R
Ivy
Cristina
C
Melton
Rachael
R
Matthews
Heather
H
White
Neill
N
Li
Feng
F
Harpin
Vanessa
V
Ecker
David J
DJ
McDougal
Linda K
LK
Limbago
Brandi
B
Ross
Tracy
T
Wolk
Donna M
DM
Wysocki
Vicki
V
Carroll
Karen C
KC
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
D017931:DNA Primers; D004269:DNA, Bacterial