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

doi:10.2217/fmb-2018-0155

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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

Journal article

Peer reviewed

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 and Derek S Sarovich

Future Microbiology, Vol.13(12), pp.1403-1418

2018

DOI: https://doi.org/10.2217/fmb-2018-0155

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Abstract

antibiotic resistance

Burkholderia pseudomallei

efflux pump

gene expression

melioidosis

qPCR

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.

Details

Title: Development and validation of a triplex quantitative real-time PCR assay to detect efflux pump-mediated antibiotic resistance in Burkholderia pseudomallei
Authors: Jessica R Webb (Author)
Erin P Price (Author) - University of the Sunshine Coast - Faculty of Science, Health, Education and Engineering
Nawarat Somprasong (Author)
Herbert P Schweizer (Author)
Robert W Baird (Author)
Bart J Currie (Author)
Derek S Sarovich (Author) - University of the Sunshine Coast - Faculty of Science, Health, Education and Engineering
Publication details: Future Microbiology, Vol.13(12), pp.1403-1418
Publisher: Future Medicine Ltd.
Date published: 2018
DOI: 10.2217/fmb-2018-0155
ISSN: 1746-0913
Organisation Unit: University of the Sunshine Coast, Queensland; Centre for Bioinnovation
Language: English
Record Identifier: 99451427002621
Output Type: Journal article

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Collaboration types: Domestic collaboration; International collaboration
Web Of Science research areas: Microbiology

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