Rapid and high-throughput genotyping of Staphylococcus epidermidis isolates by automated multilocus variable-number of tandem repeats: a tool for real-time epidemiology.
Fast and reliable genotyping methods allowing real-time epidemiology would be instrumental to discriminate Staphylococcus epidermidis isolates, in order to evaluate potential cross-infections or to follow genome content of infecting strains of this important opportunistic pathogen. We describe an automated multilocus variable-number tandem repeat-based assay (MLVA) for the rapid genotyping of S. epidermidis. Multiplex PCR amplifications using 6 primer pairs targeting gene-regions containing variable numbers of tandem repeats and the mecA gene are resolved by micro-capillary electrophoresis and automatically assessed by cluster analysis. This genotyping technique was evaluated for discriminatory power and reproducibility on 2 sequenced strains, on a collection of 21 strains previously characterized using genotyping reference methods and finally on 65 clinical isolates identified in two different institutions. All steps of this new procedure were developed to ensure rapid turn-around time and moderate costs. Our results suggest that this rapid approach is a valuable epidemiological tool to genotype S. epidermidis isolates in real-time. The rapid analysis of a limited number of evolutionary markers showed a power of discrimination similar to that of pulse-field gel electrophoresis (PFGE) or multilocus sequence type (MLST). This type of rapid and high-throughput methodology opens the possibility to rapidly assess long-term nosocomial transmission or to characterize infecting strains in the general procedure of routine laboratories, in real-time.
Francois
Patrice
P
University of Geneva Hospitals, CH-1211 Geneva 14, Switzerland. patrice.francois@genomic.ch
Hochmann
Alena
A
Huyghe
Antoine
A
Bonetti
Eve-Julie
EJ
Renzi
Gesuele
G
Harbarth
Stephan
S
Klingenberg
Claus
C
Pittet
Didier
D
Schrenzel
Jacques
J
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques / Q000379:methods*; D016000:Cluster Analysis; D004269:DNA, Bacterial / Q000235:genetics; D019075:Electrophoresis, Capillary; D005838:Genotype; D006801:Humans; D008826:Microbial Sensitivity Tests; D018598:Minisatellite Repeats; D017720:Molecular Epidemiology / Q000379:methods*; D013203:Staphylococcal Infections / Q000453:epidemiology / Q000382:microbiology*; D013212:Staphylococcus epidermidis / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004269:DNA, Bacterial; C487495:MecA protein, Staphylococcus epidermidis