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Journal of Clinical Microbiology, April 2002, p. 1333-1338, Vol. 40, No. 4
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.4.1333-1338.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

rpoB Gene Sequence-Based Identification of Staphylococcus Species

Michel Drancourt and Didier Raoult*

Unité des Rickettsies, CNRS UMR-6020, IFR 48, Faculté de Médecine, Marseille, France

Received 5 September 2001/ Returned for modification 30 October 2001/ Accepted 7 January 2002

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.

* Corresponding author. Mailing address: Unité des Rickettsies, Faculté de Médecine, 27 Boulevard Jean Moulin, 13385 Marseille cedex 5, France. Phone: 33 (0)4 91 32 43 75. Fax: 33 (0)4 91 83 03 90. E-mail: Didier.Raoult{at}medecine.univ-mrs.fr.

Journal of Clinical Microbiology, April 2002, p. 1333-1338, Vol. 40, No. 4
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.4.1333-1338.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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