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Journal of Clinical Microbiology, February 2004, p. 497-504, Vol. 42, No. 2
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.2.497-504.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

rpoB Gene Sequence-Based Identification of Aerobic Gram-Positive Cocci of the Genera Streptococcus, Enterococcus, Gemella, Abiotrophia, and Granulicatella

Michel Drancourt, Véronique Roux, Pierre-Edouard Fournier, and Didier Raoult*

Unité des Rickettsies, IFR 48, Faculté de Médecine, Université de la Méditerranée, Marseille, France

Received 28 August 2003/ Returned for modification 12 October 2003/ Accepted 27 October 2003

We developed a new molecular tool based on rpoB gene (encoding the beta subunit of RNA polymerase) sequencing to identify streptococci. We first sequenced the complete rpoB gene for Streptococcus anginosus, S. equinus, and Abiotrophia defectiva. Sequences were aligned with these of S. pyogenes, S. agalactiae, and S. pneumoniae available in GenBank. Using an in-house analysis program (SVARAP), we identified a 740-bp variable region surrounded by conserved, 20-bp zones and, by using these conserved zones as PCR primer targets, we amplified and sequenced this variable region in an additional 30 Streptococcus, Enterococcus, Gemella, Granulicatella, and Abiotrophia species. This region exhibited 71.2 to 99.3% interspecies homology. We therefore applied our identification system by PCR amplification and sequencing to a collection of 102 streptococci and 60 bacterial isolates belonging to other genera. Amplicons were obtained in streptococci and Bacillus cereus, and sequencing allowed us to make a correct identification of streptococci. Molecular signatures were determined for the discrimination of closely related species within the S. pneumoniae-S. oralis-S. mitis group and the S. agalactiae-S. difficile group. These signatures allowed us to design a S. pneumoniae-specific PCR and sequencing primer pair.

* Corresponding author. Mailing address: Unité des Rickettsies, IFR 48, Faculté de Médecine, Université de la Méditerranée, Marseille, France. Phone: 04-91-38-55-17. Fax: 04-91-38-77-72. E-mail: Didier.Raoult{at}medecine.univ-mrs.fr.

Journal of Clinical Microbiology, February 2004, p. 497-504, Vol. 42, No. 2
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.2.497-504.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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