Development of a polymerase chain reaction assay for the specific identification of Burkholderia mallei and differentiation from Burkholderia pseudomallei and other closely related Burkholderiaceae.
Burkholderia mallei and Burkholderia pseudomallei, the etiologic agents responsible for glanders and melioidosis, respectively, are genetically and phenotypically similar and are category B biothreat agents. We used an in silico approach to compare the B. mallei ATCC 23344 and B. pseudomallei K96243 genomes to identify nucleotide sequences unique to B. mallei. Five distinct B. mallei DNA sequences and/or genes were identified and evaluated for polymerase chain reaction (PCR) assay development. Genomic DNAs from a collection of 31 B. mallei and 34 B. pseudomallei isolates, obtained from various geographic, clinical, and environmental sources over a 70-year period, were tested with PCR primers targeted for each of the B. mallei ATCC 23344-specific nucleotide sequences. Of the 5 chromosomal targets analyzed, only PCR primers designed to bimA(Bm) were specific for B. mallei. These primers were used to develop a rapid PCR assay for the definitive identification of B. mallei and differentiation from all other bacteria.
Ulrich
Ricky L
RL
Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD 21702-5011, USA.
Ulrich
Melanie P
MP
Schell
Mark A
MA
Kim
H Stanley
HS
DeShazer
David
D
D015373:Bacterial Typing Techniques / Q000379:methods*; D042726:Burkholderia mallei / Q000235:genetics* / Q000302:isolation & purification; D016957:Burkholderia pseudomallei / Q000235:genetics* / Q000302:isolation & purification; D017931:DNA Primers / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics*; D005798:Genes, Bacterial; D005838:Genotype; D005896:Glanders / Q000175:diagnosis / Q000382:microbiology; D006801:Humans; D008554:Melioidosis / Q000175:diagnosis / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004269:DNA, Bacterial