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

Jessica R. Webb, Erin P. Price, Nawarat Somprasong, Herbert P. Schweizer, Robert W. Baird, Bart J. Currie, Derek S. Sarovich

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)1403-1418
Number of pages16
JournalFuture Microbiology
Volume13
Issue number12
DOIs
StatePublished - Sep 2018
Externally publishedYes

Keywords

  • Burkholderia pseudomallei
  • antibiotic resistance
  • efflux pump
  • gene expression
  • melioidosis
  • qPCR

ASJC Scopus subject areas

  • Microbiology
  • Microbiology (medical)

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