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Applied and Environmental Microbiology, January 2004, p. 191-201, Vol. 70, No. 1
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.1.191-201.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Multilocus Sequence Typing Scheme for Bacteria of the Bacillus cereus Group

Erlendur Helgason,1 Nicolas J. Tourasse,1 Roger Meisal,1 Dominique A. Caugant,2,3 and Anne-Brit Kolstø1*

Biotechnology Centre of Oslo and School of Pharmacy,1 Institute of Oral Biology, Dental Faculty, University of Oslo,3 Division of Infectious Disease Control, Norwegian Institute of Public Health, Oslo, Norway2

Received 23 June 2003/ Accepted 7 October 2003

In this study we developed a multilocus sequence typing (MLST) scheme for bacteria of the Bacillus cereus group. This group, which includes the species B. cereus, B. thuringiensis, B. weihenstephanensis, and B. anthracis, is known to be genetically very diverse. It is also very important because it comprises pathogenic organisms as well as bacteria with industrial applications. The MLST system was established by using 77 strains having various origins, including humans, animals, food, and soil. A total of 67 of these strains had been analyzed previously by multilocus enzyme electrophoresis, and they were selected to represent the genetic diversity of this group of bacteria. Primers were designed for conserved regions of housekeeping genes, and 330- to 504-bp internal fragments of seven such genes, adk, ccpA, ftsA, glpT, pyrE, recF, and sucC, were sequenced for all strains. The number of alleles at individual loci ranged from 25 to 40, and a total of 53 allelic profiles or sequence types (STs) were distinguished. Analysis of the sequence data showed that the population structure of the B. cereus group is weakly clonal. In particular, all five B. anthracis isolates analyzed had the same ST. The MLST scheme which we developed has a high level of resolution and should be an excellent tool for studying the population structure and epidemiology of the B. cereus group.

* Corresponding author. Mailing address: Biotechnology Centre of Oslo, University of Oslo, P.O. Box 1125, 0316 Oslo, Norway. Phone: 47 22845677. Fax: 47 22840501. E-mail: annebko{at}biotek.uio.no.

Applied and Environmental Microbiology, January 2004, p. 191-201, Vol. 70, No. 1
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.1.191-201.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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