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Journal of Clinical Microbiology, October 2003, p. 4647-4654, Vol. 41, No. 10
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.10.4647-4654.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Use of 16S rRNA Gene Sequencing for Rapid Identification and Differentiation of Burkholderia pseudomallei and B. mallei

Jay E. Gee,1* Claudio T. Sacchi,1,2 Mindy B. Glass,1 Barun K. De,1 Robbin S. Weyant,3 Paul N. Levett,1 Anne M. Whitney,1 Alex R. Hoffmaster,1 and Tanja Popovic1

Meningitis and Special Pathogens Branch, Division of Bacterial and Mycotic Diseases, National Center for Infectious Diseases,1 Office of Health and Safety, Centers for Disease Control and Prevention, Atlanta, Georgia 30333,3 Adolfo Lutz Institute, São Paulo, Brazil2

Received 19 May 2003/ Returned for modification 7 July 2003/ Accepted 14 July 2003

Burkholderia pseudomallei and B. mallei, the causative agents of melioidosis and glanders, respectively, are designated category B biothreat agents. Current methods for identifying these organisms rely on their phenotypic characteristics and an extensive set of biochemical reactions. We evaluated the use of 16S rRNA gene sequencing to rapidly identify these two species and differentiate them from each other as well as from closely related species and genera such as Pandoraea spp., Ralstonia spp., Burkholderia gladioli, Burkholderia cepacia, Burkholderia thailandensis, and Pseudomonas aeruginosa. We sequenced the 1.5-kb 16S rRNA gene of 56 B. pseudomallei and 23 B. mallei isolates selected to represent a wide range of temporal, geographic, and origin diversity. Among all 79 isolates, a total of 11 16S types were found based on eight positions of difference. Nine 16S types were identified in B. pseudomallei isolates based on six positions of difference, with differences ranging from 0.5 to 1.5 bp. Twenty-two of 23 B. mallei isolates showed 16S rRNA gene sequence identity and were designated 16S type 10, whereas the remaining isolate was designated type 11. This report provides a basis for rapidly identifying and differentiating B. pseudomallei and B. mallei by molecular methods.

* Corresponding author. Mailing address: Meningitis and Special Pathogens Branch, Division of Bacterial and Mycotic Diseases, National Center for Infectious Diseases, CDC, MS-D11, 1600 Clifton Rd., N.E., Atlanta, GA 30333. Phone: (404) 639-4936. Fax: (404) 639-4421. E-mail: JGee1{at}cdc.gov.

Journal of Clinical Microbiology, October 2003, p. 4647-4654, Vol. 41, No. 10
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.10.4647-4654.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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