title: Use of 16S rRNA gene sequencing for rapid identification and differentiation of Burkholderia pseudomallei and B. mallei.
abstract: 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.
authors.affiliation: Meningitis and Special Pathogens Branch, Division of Bacterial and Mycotic Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA. JGee1@cdc.gov
authors.forename: Jay E
authors.initials: JE
authors.lastname: Gee
authors.forename: Claudio T
authors.initials: CT
authors.lastname: Sacchi
authors.forename: Mindy B
authors.initials: MB
authors.lastname: Glass
authors.forename: Barun K
authors.initials: BK
authors.lastname: De
authors.forename: Robbin S
authors.initials: RS
authors.lastname: Weyant
authors.forename: Paul N
authors.initials: PN
authors.lastname: Levett
authors.forename: Anne M
authors.initials: AM
authors.lastname: Whitney
authors.forename: Alex R
authors.initials: AR
authors.lastname: Hoffmaster
authors.forename: Tanja
authors.initials: T
authors.lastname: Popovic
mesh_terms: D000818:Animals; D015373:Bacterial Typing Techniques; D019117:Burkholderia / Q000145:classification* / Q000235:genetics; D019121:Burkholderia Infections / Q000382:microbiology; D016957:Burkholderia pseudomallei / Q000145:classification* / Q000235:genetics; D004275:DNA, Ribosomal / Q000032:analysis; D004783:Environmental Microbiology; D020459:Genes, rRNA; D006801:Humans; D008554:Melioidosis / Q000382:microbiology; D008969:Molecular Sequence Data; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D017422:Sequence Analysis, DNA
chemical_list: D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S