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Journal of Clinical Microbiology, April 2004, p. 1626-1630, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1626-1630.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Reassessment of Sequence-Based Targets for Identification of Bacillus Species

K. S. Blackwood,^1* C. Y. Turenne,¹ D. Harmsen,² and A. M. Kabani^1,3

National Reference Centre for Mycobacteriology, National Microbiology Laboratory, Population and Public Health Branch, Health Canada,¹ Department of Medical Microbiology, University of Manitoba, Winnipeg, Manitoba, Canada,³ Institut für Hygiene und Mikrobiologie, Universität Münster, Münster, Germany²

Received 13 August 2003/ Returned for modification 7 October 2003/ Accepted 14 December 2003

The Bacillus genus is a large heterogeneous group in need of an efficient method for species differentiation. To determine the current validity of a sequence-based method for identification and provide contemporary data, PCR and sequencing of a 500-bp product encompassing the V1 to V3 regions of the 16S rRNA gene were undertaken using 65 of the 83 type strains of this genus. This region proved discriminatory between most species (70.0 to 100% similarity), the exceptions being clinically relevant B. cereus and B. anthracis as well as nonpathogenic B. psychrotolerans and B. psychrodurans. Consequently, 27 type and clinical strains from the B. cereus group were used to test alternate targets (rpoB, vrrA, and the 16S-23S spacer region) for identification. The rpoB gene proved the best alternate target, with a conserved 4-nucleotide difference between B. cereus and B. anthracis. The high 16S rRNA gene sequence similarities between some strains demonstrated the need for a polyphasic approach to the systematics of this genus. This approach is one focus of the Ribosomal Differentiation of Medical Microorganisms mandate. Accordingly, the 16S rRNA gene sequences generated in this study have been submitted for inclusion into its publicly accessible, quality-controlled database at http://www.ridom_rdna.de/.

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* Corresponding author. Mailing address: National Reference Centre for Mycobacteriology, Canadian Science Centre for Human and Animal Health, 1015 Arlington St., Winnipeg, Manitoba, Canada R3E 3R2. Phone: (204) 789-6039. Fax: (204) 789-2036. E-mail: kym_blackwood{at}hc-sc.gc.ca.

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Journal of Clinical Microbiology, April 2004, p. 1626-1630, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1626-1630.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

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