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Journal of Clinical Microbiology, February 2006, p. 536-546, Vol. 44, No. 2
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.2.536-546.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Use of PCR-Restriction Enzyme Pattern Analysis and Sequencing Database for hsp65 Gene-Based Identification of Nocardia Species

Verónica Rodríguez-Nava,1 Andrée Couble,1 Gregory Devulder,2 Jean-Pierre Flandrois,2 Patrick Boiron,1 and Frédéric Laurent1*

UMR CNRS 5557, Center for Microbial Ecology, Opportunistic Pathogens and Environmental Research Group, French Observatory for Nocardiosis, Laboratoire de Mycologie Fondamentale et Appliquée aux Biotechnologies Industrielles, UFR de Pharmacie, Université Claude Bernard Lyon I, Lyon, France,1 UMR CNRS 5558, Laboratoire de Bactériologie, UFR de Médecine Lyon-Sud, BP 12, 69921 Oullins Cedex, France2

Received 6 October 2004/ Returned for modification 30 November 2004/ Accepted 22 November 2005

Nocardia identification required laborious and time-consuming phenotypic and chemotaxonomic methods until molecular methods were developed in the mid-1990s. Here we reassessed the capacity of PCR-restriction enzyme pattern analysis (PRA) of the hsp65 gene to differentiate Nocardia species, including 36 new species. Our results confirm that hsp65 PRA must no longer be used for Nocardia species identification, as many species have the same restriction pattern. We then compared sequencing-based strategies using an hsp65 database and a 16S rRNA database and found that the hsp65 region contained sufficient polymorphisms for comprehensive Nocardia species identification.

* Corresponding author. Mailing address: UMR CNRS 5557, Laboratoire de Mycologie Fondamentale et Appliquée aux Biotechnologies Industrielles, UFR Pharmacie, UCB Lyon I, 69373, Lyon Cedex 08, France. Phone: (33) (0) 4 78 77 70 00. Fax: (33) (0) 4 78 77 72 12. E-mail: flaurent{at}ch-bourg01.fr.

Journal of Clinical Microbiology, February 2006, p. 536-546, Vol. 44, No. 2
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.2.536-546.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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