title: rpoB gene sequence-based identification of Staphylococcus species.
abstract: The complete sequence of rpoB, the gene encoding the beta subunit of RNA polymerase was determined for Staphylococcus saccharolyticus, Staphylococcus lugdunensis, S taphylococcus caprae, and Staphylococcus intermedius and partial sequences were obtained for an additional 27 Staphylococcus species. The complete rpoB sequences varied in length from 3,452 to 3,845 bp and had a 36.8 to 39.2% GC content. The partial sequences had 71.6 to 93.6% interspecies homology and exhibited a 0.08 to 0.8% intraspecific divergence. With a few exceptions, the phylogenetic relationships inferred from the partial rpoB sequences were in agreement with those previously derived from DNA-DNA hybridization studies and analyses of 16S ribosomal DNA gene sequences and partial HSP60 gene sequences. The staphylococcal rpoB sequence database we established enabled us to develop a molecular method for identifying Staphylococcus isolates by PCR followed by direct sequencing of the 751-bp amplicon. In blind tests, this method correctly identified 10 Staphylococcus isolates, and no positive results were obtained with 10 non-Staphylococcus gram-positive and gram-negative bacterial isolates. We propose partial sequencing of the rpoB gene as a new tool for the accurate identification of Staphylococcus isolates.
authors.affiliation: Unité des Rickettsies, CNRS UMR-6020, IFR 48, Faculté de Médecine, Marseille, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Drancourt
authors.forename: Didier
authors.initials: D
authors.lastname: Raoult
mesh_terms: D015373:Bacterial Typing Techniques; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D016208:Databases, Factual; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013210:Staphylococcus / Q000145:classification* / Q000235:genetics
chemical_list: D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit