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Journal of Clinical Microbiology, July 2003, p. 2908-2914, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.2908-2914.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Identification of Bacillus anthracis by rpoB Sequence Analysis and Multiplex PCR

Kwan Soo Ko,1 Jong-Man Kim,2 Jong-Wan Kim,2 Byeong Yeal Jung,3 Wonyong Kim,4 Ik Jung Kim,5 and Yoon-Hoh Kook1*

Department of Microbiology and Cancer Research Institute, Institute of Endemic Diseases, SNUMRC, Seoul National University College of Medicine, and Clinical Research Institute, Seoul National University Hospital, Seoul 110-799,1 Animal Disease Research Department,2 Department of Pharmacology, National Veterinary Research and Quarantine Service, Kyunggi Province, 430-824,3 Department of Microbiology, Chung-Ang University College of Medicine and Institute of Medical Research, Seoul 156-756,4 Department of Microbiology, School of Medicine, Dongguk University, Gyungju, Korea5

Received 13 January 2003/ Returned for modification 18 February 2003/ Accepted 4 April 2003

Comparative sequence analysis was performed upon Bacillus anthracis and its closest relatives, B. cereus and B. thuringiensis. Portions of rpoB DNA from 10 strains of B. anthracis, 16 of B. cereus, 10 of B. thuringiensis, 1 of B. mycoides, and 1 of B. megaterium were amplified and sequenced. The determined rpoB sequences (318 bp) of the 10 B. anthracis strains, including five Korean isolates, were identical to those of Ames, Florida, Kruger B, and Western NA strains. Strains of the "B. cereus group" were separated into two subgroups, in which the B. anthracis strains formed a separate clade in the phylogenetic tree. However, B. cereus and B. thuringiensis could not be differentiated. Sequence analysis confirmed the five Korean isolates as B. anthracis. Based on the rpoB sequences determined in the present study, multiplex PCR generating either B. anthracis-specific amplicons (359 and 208 bp) or cap DNA (291 bp) in a virulence plasmid could be used for the rapid differential detection and identification of virulent B. anthracis.

* Corresponding author. Mailing address: Department of Microbiology, Seoul National University College of Medicine, 28 Yongon-dong, Chongno-gu, Seoul 110-799, Korea. Phone: (82) 2-740-8306. Fax: (82) 2-743-0881. E-mail: yhkook{at}plaza.snu.ac.kr.

Journal of Clinical Microbiology, July 2003, p. 2908-2914, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.2908-2914.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Valjevac, S., Hilaire, V., Lisanti, O., Ramisse, F., Hernandez, E., Cavallo, J.-D., Pourcel, C., Vergnaud, G. (2005). Comparison of Minisatellite Polymorphisms in the Bacillus cereus Complex: a Simple Assay for Large-Scale Screening and Identification of Strains Most Closely Related to Bacillus anthracis. Appl. Environ. Microbiol. 71: 6613-6623 [Abstract] [Full Text]  
  • Ko, K. S., Kim, J.-W., Kim, J.-M., Kim, W., Chung, S.-i., Kim, I. J., Kook, Y.-H. (2004). Population Structure of the Bacillus cereus Group as Determined by Sequence Analysis of Six Housekeeping Genes and the plcR Gene. Infect. Immun. 72: 5253-5261 [Abstract] [Full Text]  
  • Korczak, B., Christensen, H., Emler, S., Frey, J., Kuhnert, P. (2004). Phylogeny of the family Pasteurellaceae based on rpoB sequences. Int. J. Syst. Evol. Microbiol. 54: 1393-1399 [Abstract] [Full Text]  


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