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Journal of Clinical Microbiology, January 2004, p. 179-185, Vol. 42, No. 1
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.1.179-185.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Detection of the Bacillus anthracis gyrA Gene by Using a Minor Groove Binder Probe

William Hurtle,1 Elizabeth Bode,2 David A. Kulesh,2 Rebecca Susan Kaplan,2 Jeff Garrison,3 Deanna Bridge,2 Michelle House,2 Melissa S. Frye,4 Bonnie Loveless,4 and David Norwood2*

Clinical Research Management, North Royalton, Ohio 44133,1 U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland 21701-5011,2 Battelle, Columbus, Ohio 43201,3 Geo-Centers, Newtown, Massachusetts 024594

Received 23 May 2003/ Returned for modification 8 July 2003/ Accepted 26 August 2003

Identification of chromosomal markers for rapid detection of Bacillus anthracis is difficult because significant chromosomal homology exists among B. anthracis, Bacillus cereus, and Bacillus thuringiensis. We evaluated the bacterial gyrA gene as a potential chromosomal marker for B. anthracis. A real-time PCR assay was developed for the detection of B. anthracis. After analysis of the unique nucleotide sequence of the B. anthracis gyrA gene, a fluorescent 3' minor groove binding probe was tested with 171 organisms from 29 genera of bacteria, including 102 Bacillus strains. The assay was found to be specific for all 43 strains of B. anthracis tested. In addition, a test panel of 105 samples was analyzed to evaluate the potential diagnostic capability of the assay. The assay showed 100% specificity, demonstrating the usefulness of the gyrA gene as a specific chromosomal marker for B. anthracis.

* Corresponding author. Mailing address: U.S. Army Medical Research Institute of Infectious Diseases, Diagnostic Systems Division, 1425 Porter St., Fort Detrick, Frederick, MD 21702. Phone: (301) 619-4721. Fax: (301) 619-2492. E-mail: David.Norwood{at}det.amedd.army.mil.

Journal of Clinical Microbiology, January 2004, p. 179-185, Vol. 42, No. 1
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.1.179-185.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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