Application of Multilocus Sequence Analysis (MLSA) For Rapid Identification of Enterococcus Species Based on Rpoa And Phes Genes

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Journal Title, Volume, Page: 
Microbiology 151, 2141-215
Year of Publication: 
2005
Authors: 
Sabri M. Naser
Laboratory of Microbiology, Ghent University, K. L. Ledeganckstraat 35, Ghent 9000, Belgium
Current Affiliation: 
Department of Biology, Faculty of Science, An-Najah National University, Nablus. Palestine
Fabiano L. Thompson
BCCM/LMG Bacteria Collection, Ghent University, K. L. Ledeganckstraat 35, Ghent 9000, Belgium
Bart Hoste
BCCM/LMG Bacteria Collection, Ghent University, K. L. Ledeganckstraat 35, Ghent 9000, Belgium
Dirk Gevers
BCCM/LMG Bacteria Collection, Ghent University, K. L. Ledeganckstraat 35, Ghent 9000, Belgium
Peter Dawyndt
Laboratory of Microbiology, Ghent University, K. L. Ledeganckstraat 35, Ghent 9000, Belgium
Marc Vancanneyt
BCCM/LMG Bacteria Collection, Ghent University, K. L. Ledeganckstraat 35, Ghent 9000, Belgium
Jean Swings
BCCM/LMG Bacteria Collection, Ghent University, K. L. Ledeganckstraat 35, Ghent 9000, Belgium
Preferred Abstract (Original): 
The aim of our study was to evaluate the use of RNA polymerase alpha subunit (rpoA) and phenylalanyl-tRNA synthase (pheS) gene sequences as species identification tools for enterococci. Ninety-six representative strains comprising all currently recognized Enterococcus species were examined. rpoA gene sequences generated a robust classification into species groups similar to the one based on the 16S rRNA gene sequence analysis. On the other hand, pheS gene is a fast evolving clock even better suited for species delineation than rpoA gene, but not for recognition of the species groups within Enterococcus as determined by both rpoA and 16S rRNA genes. All enterococcal species were clearly differentiated on the basis of rpoA and pheS sequences. Evaluation of the intraspecies variation showed that both rpoA and pheS genes had a high degree of homogeneity among strains of the same species. Strains of the same enterococcal species have at least 99 % rpoA and 97 % pheS gene sequence similarity, whereas, different enterococcal species have at maximum 97 % rpoA and 86 % pheS gene sequence similarity. We can conclude that both genes can be used as reliable tools for identification of clinical and environmental species of Enterococcus and are efficient screening methods for detection of novel species. The sequence data obtained in this study was compared to the available atpA and 16S rRNA gene sequences. The MLSA approach of Enterococcus provides portable, highly reproducible data with lower costs for rapid identification of all enterococcal species.
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