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 journalArticleResearchpeer-review

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
Pages (from-to)1403-1418
Number of pages16
JournalFuture Microbiology
Volume13
Issue number12
Early online date26 Sep 2018
DOIs
Publication statusPublished - Sep 2018

Fingerprint

Burkholderia pseudomallei
Multiplex Polymerase Chain Reaction
meropenem
Microbial Drug Resistance
Real-Time Polymerase Chain Reaction
Up-Regulation
Doxycycline
Sulfamethoxazole Drug Combination Trimethoprim
Microbial Sensitivity Tests
Treatment Failure
RNA
Anti-Bacterial Agents
Mutation
Infection
Therapeutics

Cite this

Webb, Jessica R. ; Price, Erin P. ; Somprasong, Nawarat ; Schweizer, Herbert P. ; Baird, Robert W. ; Currie, Bart J. ; Sarovich, Derek S. / Development and validation of a triplex quantitative real-time PCR assay to detect efflux pump-mediated antibiotic resistance in Burkholderia pseudomallei. In: Future Microbiology. 2018 ; Vol. 13, No. 12. pp. 1403-1418.
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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.",
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Development and validation of a triplex quantitative real-time PCR assay to detect efflux pump-mediated antibiotic resistance in Burkholderia pseudomallei. / Webb, Jessica R.; Price, Erin P.; Somprasong, Nawarat; Schweizer, Herbert P.; Baird, Robert W.; Currie, Bart J.; Sarovich, Derek S.

In: Future Microbiology, Vol. 13, No. 12, 09.2018, p. 1403-1418.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Webb, Jessica R.

AU - Price, Erin P.

AU - Somprasong, Nawarat

AU - Schweizer, Herbert P.

AU - Baird, Robert W.

AU - Currie, Bart J.

AU - Sarovich, Derek S.

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AB - 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.

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