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Applied and Environmental Microbiology, July 2001, p. 3021-3028, Vol. 67, No. 7
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.7.3021-3028.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Use of Long-Range Repetitive Element Polymorphism-PCR To Differentiate Bacillus anthracis Strains

Michael J. Brumlik, Urszula Szymajda,dagger Dorota Zakowska, Xudong Liang, Rajendra J. Redkar, Guy Patra, and Vito G. Del Vecchio*

Institute of Molecular Biology and Medicine, University of Scranton, Scranton, Pennsylvania 18510

Received 20 November 2000/Accepted 11 April 2001

The genome of Bacillus anthracis is extremely monomorphic, and thus individual strains have often proven to be recalcitrant to differentiation at the molecular level. Long-range repetitive element polymorphism-PCR (LR REP-PCR) was used to differentiate various B. anthracis strains. A single PCR primer derived from a repetitive DNA element was able to amplify variable segments of a bacterial genome as large as 10 kb. We were able to characterize five genetically distinct groups by examining 105 B. anthracis strains of diverse geographical origins. All B. anthracis strains produced fingerprints comprising seven to eight bands, referred to as "skeleton" bands, while one to three "diagnostic" bands differentiated between B. anthracis strains. LR REP-PCR fingerprints of B. anthracis strains showed very little in common with those of other closely related species such as B. cereus, B. thuringiensis, and B. mycoides, suggesting relative heterogeneity among the non-B. anthracis strains. Fingerprints from transitional non-B. anthracis strains, which possessed the B. anthracis chromosomal marker Ba813, scarcely resembled those observed for any of the five distinct B. anthracis groups that we have identified. The LR REP-PCR method described in this report provides a simple means of differentiating B. anthracis strains.

* Corresponding author. Mailing address: Institute of Molecular Biology and Medicine, University of Scranton, Scranton PA 18510-4625. Phone: (570) 941-4817. Fax: (570) 941-6229. E-mail: vimbm{at}aol.com.

dagger Present address: The Military Institute of Hygiene and Epidemiology, Pulawy, Poland.

Applied and Environmental Microbiology, July 2001, p. 3021-3028, Vol. 67, No. 7
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.7.3021-3028.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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