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Journal of Clinical Microbiology, February 2003, p. 657-666, Vol. 41, No. 2
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.2.657-666.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Identification of Major Streptococcal Species by rrn-Amplified Ribosomal DNA Restriction Analysis

Laurent Schlegel,1,2 Francine Grimont,2 Patrick A. D. Grimont,2 and Anne Bouvet1*

Centre National de Référence des Streptocoques, Service de Microbiologie, Hôtel Dieu, AP-HP Université Paris VI,1 Unité Biodiversité des Bactéries Pathogènes Emergentes, INSERM U389, Institut Pasteur, Paris, France2

Received 20 May 2002/ Returned for modification 6 August 2002/ Accepted 31 October 2002

Amplified ribosomal DNA restriction analysis (rrn-ARDRA) is based on PCR amplification and restriction of a fragment of rRNA genes including 16S and 23S genes and the intergenic spacer. rrn-ARDRA was evaluated for the identification of species within the genus Streptococcus. A total of 148 type and reference strains of pyogenic, oral, and group D streptococci were examined in order to construct a database for identification of streptococci. The amplified product was a single band approximately 4,500 bp long. This amplicon was digested separately with three (HhaI, MboII, and Sau3A) restriction endonucleases. Respectively, 27, 26, and 28 major patterns were observed after HhaI, MboII, and Sau3A restrictions. Streptococcal strains belonging to different species had different patterns or different combination of patterns. An identification system based upon a combination of the three restriction patterns in a single database was then proposed. rrn-ARDRA was successfully applied to 11 clinical isolates whose identification to the species level was difficult to obtain by phenotypic analysis. Using a database of well-characterized strains, rrn-ARDRA is a powerful method for the identification of streptococcal isolates.

* Corresponding author. Mailing address: Centre National de Référence des Streptocoques, Service de Microbiologie, Hôtel Dieu, Université Paris VI, 1, place du Parvis Notre-Dame, F-75181 Paris cedex 04, France. Phone: 33 1 42 34 82 73. Fax: 33 1 42 34 87 19. E-mail: anne.bouvet{at}htd.ap-hop-paris.fr.

Journal of Clinical Microbiology, February 2003, p. 657-666, Vol. 41, No. 2
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.2.657-666.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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