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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
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- Title
- Development and validation of a triplex quantitative real-time PCR assay to detect efflux pump-mediated antibiotic resistance in Burkholderia pseudomallei
- Author/Creator
-
Webb, Jessica R |
Price, Erin P |
Somprasong, Nawarat |
Schweizer, Herbert P |
Baird, Robert W |
Currie, Bart J |
Sarovich, D S
- Description
- 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.
- Relation
- Future Microbiology / Vol. 13, No. 12, pp.1403-1418
- Relation
- http://dx.doi.org/10.2217/fmb-2018-0155
- Year
- 2018
- Publisher
- Future Medicine Ltd.
- Subject
-
FoR 0605 (Microbiology) |
FoR 1108 (Medical Microbiology) |
antibiotic resistance |
Burkholderia pseudomallei |
efflux pump |
gene expression |
melioidosis |
qPCR
- Resource Type
- Journal Article
- Identifier
- ISSN: 1746-0913
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