Use of an automated multiple-locus, variable-number tandem repeat-based method for rapid and high-throughput genotyping of Staphylococcus aureus isolates.
Fast and reliable genotyping methods that allow real-time epidemiological surveillance would be instrumental to monitoring of the spread of methicillin-resistant Staphylococcus aureus. We describe an automated variable-number tandem repeat-based method for the rapid genotyping of Staphylococcus aureus. Multiplex PCR amplifications with eight primer pairs that target gene regions with variable numbers of tandem repeats were resolved by microcapillary electrophoresis and automatically assessed by cluster analysis. This genotyping technique was evaluated for its discriminatory power and reproducibility with clinical isolates of various origins, including a panel of control strains previously characterized by several typing methods and collections from either long-term carriers or defined nosocomial outbreaks. All steps of this new procedure were developed to ensure a rapid turnaround time and moderate cost. The results obtained suggest that this rapid approach is a valuable tool for the genotyping of S. aureus isolates in real time.
Genomic Research Laboratory, University of Geneva Hospitals, CH-1211 Geneva 14, Switzerland. patrice.francois@genomic.ch
Patrice
P
Francois
Antoine
A
Huyghe
Yvan
Y
Charbonnier
Manuela
M
Bento
Sébastien
S
Herzig
Ivan
I
Topolski
Bénédicte
B
Fleury
Daniel
D
Lew
Pierre
P
Vaudaux
Stephan
S
Harbarth
Willem
W
van Leeuwen
Alex
A
van Belkum
Dominique S
DS
Blanc
Didier
D
Pittet
Jacques
J
Schrenzel
D001331:Automation; D001426:Bacterial Proteins / Q000235:genetics*; D015373:Bacterial Typing Techniques; D004196:Disease Outbreaks; D016521:Electrophoresis, Gel, Pulsed-Field; D005838:Genotype; D006801:Humans; D016106:Methicillin Resistance; D018598:Minisatellite Repeats / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D013203:Staphylococcal Infections / Q000453:epidemiology / Q000382:microbiology; D013211:Staphylococcus aureus / Q000145:classification* / Q000187:drug effects / Q000235:genetics; D013997:Time Factors
D001426:Bacterial Proteins