Development and validation of a triplex quantitative real-time PCR assay to detect efflux pump-mediated antibiotic resistance in Burkholderia pseudomallei. (September 2018)

• Record Type: Journal Article
• Authors: Webb, Jessica R; Price, Erin P; Somprasong, Nawarat; Schweizer, Herbert P; Baird, Robert W; Currie, Bart J; Sarovich, Derek S
• Is Part Of: Future microbiology. Volume 13:Number 12(2018)
• Page Start: 1403
• Page End: 1418
• Publication Date: 2018-09
• Subjects: antibiotic resistance -- Burkholderia pseudomallei -- efflux pump -- gene expression -- melioidosis -- qPCR; Microbiology -- Research -- Periodicals; 616.9041
• DOI: 10.2217/fmb-2018-0155
• Languages: English
• ISSNs: 1746-0913

Abstract

Aim: To develop a probe-based triplex quantitative real-time PCR assay to simultaneously detect the upregulation of the efflux pumps AmrAB–OprA, BpeAB–OprB and BpeEF–OprC in Burkholderia pseudomallei strains exhibiting increased minimum inhibitory concentrations toward meropenem, doxycycline or trimethoprim-sulfamethoxazole.Methods: The triplex assay was developed and subsequently tested on RNA isolated from eight clinical and eight laboratory-generated B. pseudomallei mutants harboring efflux pump regulator mutations.Results: The triplex assay accurately detected efflux pump upregulation in all clinical and laboratory mutants, which corresponded with decreased antibiotic susceptibility or antibiotic resistance.Conclusion: Rapid detection of antibiotic resistance provides clinicians with a tool to identify potential treatment failure in near real time, enabling informed alteration of treatment during an infection and improved patient outcomes.