Use of 16S rRNA Gene Sequencing for Rapid Identification and Differentiation ofBurkholderia pseudomalleiandB. mallei

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Abstract
Burkholderia pseudomalleiandB. 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 asPandoraeaspp.,Ralstoniaspp.,Burkholderia gladioli,Burkholderia cepacia,Burkholderia thailandensis, andPseudomonas aeruginosa. We sequenced the 1.5-kb 16S rRNA gene of 56B. pseudomalleiand 23B. malleiisolates 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 inB. pseudomalleiisolates based on six positions of difference, with differences ranging from 0.5 to 1.5 bp. Twenty-two of 23B. malleiisolates 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 differentiatingB. pseudomalleiandB. malleiby molecular methods.

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