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Journal of Clinical Microbiology, September 2004, p. 3925-3931, Vol. 42, No. 9
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.9.3925-3931.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

rpoB Gene Sequencing for Identification of Corynebacterium Species

Atieh Khamis, Didier Raoult, and Bernard La Scola*

Unité des Rickettsies, CNRS UMR 6020, IFR 48, Faculté de Médecine, Université de la Méditerrannée, Marseille, France

Received 5 December 2003/ Returned for modification 12 January 2004/ Accepted 20 January 2004

The genus Corynebacterium is a heterogeneous group of species comprising human and animal pathogens and environmental bacteria. It is defined on the basis of several phenotypic characters and the results of DNA-DNA relatedness and, more recently, 16S rRNA gene sequencing. However, the 16S rRNA gene is not polymorphic enough to ensure reliable phylogenetic studies and needs to be completely sequenced for accurate identification. The almost complete rpoB sequences of 56 Corynebacterium species were determined by both PCR and genome walking methods. In all cases the percent similarities between different species were lower than those observed by 16S rRNA gene sequencing, even for those species with degrees of high similarity. Several clusters supported by high bootstrap values were identified. In order to propose a method for strain identification which does not require sequencing of the complete rpoB sequence (approximately 3,500 bp), we identified an area with a high degree of polymorphism, bordered by conserved sequences that can be used as universal primers for PCR amplification and sequencing. The sequence of this fragment (434 to 452 bp) allows accurate species identification and may be used in the future for routine sequence-based identification of Corynebacterium species.

* Corresponding author. Mailing address: Unité des Rickettsies, CNRS UMR 6020, IFR 48, Faculté de Médecine, Université de la Méditerrannée, 27 Boulevard Jean Moulin, 13385 Marseille Cedex 05, France. Phone: 33.04.91.38.55.17. Fax: 33.04.91.83.03.90. E-mail: bernard.lascola{at}medecine.univ-mrs.fr.

Journal of Clinical Microbiology, September 2004, p. 3925-3931, Vol. 42, No. 9
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.9.3925-3931.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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