Application of multilocus sequence analysis (MLSA) for rapid identification of Enterococcus species based on rpoA and pheS genes

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Abstract
The aim of this study was to evaluate the use of RNA polymeraseαsubunit (rpoA) and phenylalanyl-tRNA synthase (pheS) gene sequences as species identification tools for enterococci. Ninety-six representative strains comprising all currently recognizedEnterococcusspecies were examined.rpoAgene sequences generated a robust classification into species groups similar to the one based on 16S rRNA gene sequence analysis. On the other hand, thepheSgene is a fast-evolving clock even better suited for species delineation than therpoAgene, but not for recognition of species groups withinEnterococcusas determined by bothrpoAand 16S rRNA genes. All enterococcal species were clearly differentiated on the basis of theirrpoAandpheSsequences. Evaluation of intraspecies variation showed that bothrpoAandpheSgenes have a high degree of homogeneity among strains of the same species. Strains of the same enterococcal species have at least 99 %rpoAand 97 %pheSgene sequence similarity, whereas, different enterococcal species have at maximum 97 %rpoAand 86 %pheSgene sequence similarity. It was concluded that both genes can be used as reliable tools for identification of clinical and environmental species ofEnterococcusand are efficient screening methods for the detection of novel species. The sequence data obtained in this study were compared to the availableatpAand 16S rRNA gene sequences. The MLSA approach toEnterococcustaxonomy provides portable, highly reproducible data with lower costs for rapid identification of all enterococcal species.

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