A Quadruplex Real-Time PCR Assay for the Rapid Detection and Differentiation of the Most Relevant Members of the B. pseudomallei Complex: B. mallei, B. pseudomallei, and B. thailandensis

• DOI: 10.1371/journal.pone.0164006
• Publication Date: 2016-10-13T18:02:23Z
• Authors: Chinn-Woan Lowe; Benjamin A. Satte...; Daniel B. Nelson; Joseph D. Thiriot; Michael J. Heder; Jordon K. March; David S. Drake; Cynthia S. Lew; Annette J. Bunnell; Emily S. Moore; Kim L. O'Neill; Richard A. Robison
• Keywords: Burkholderia pseudomallei; Melioidosis

The Burkholderia pseudomallei complex classically consisted of B. mallei, pseudomallei, and thailandensis, but has now expanded to include oklahomensis, humptydooensis, three unassigned clades. Methods for detecting differentiating the been topic recent research due phenotypic genotypic similarities these species. mallei are recognized as CDC Tier 1 select agents, causative agents glanders melioidosis, respectively. Although thailandensis oklahomensis generally avirulent, both display similar characteristics that pseudomallei. humptydooensis clades genetically complex, not associated with disease. Optimal identification species remains problematic, PCR-based methods can resolve issues detection differentiation. Currently, no PCR assay is available detects major complex. A real-time in a multiplex single-tube format was developed simultaneously detect differentiate common sequence found oklahomensis. total 309 isolates 5 other bacterial were evaluated. 100% sensitive specific, demonstrated sensitivity beyond culture GC tested, completed about an hour limit between 2.6pg 48.9pg gDNA. Bioinformatic analyses also showed likely specific all 84 fully sequenced strains currently GenBank. For reasons, this could be rapid tool differentiation those clinical practical significance.