Please use this identifier to cite or link to this item: http://docs.prosentient.com.au/prosentientjspui/handle/1/11281
Title: Development and validation of a triplex quantitative real-time PCR assay to detect efflux pump-mediated antibiotic resistance in Burkholderia pseudomallei.
Authors: Webb, Jessica R
Price, Erin P
Somprasong, Nawarat
Schweizer, Herbert P
Baird, Robert W
Currie, Bart J
Sarovich, Derek S
Affiliation: Global & Tropical Health Division, Menzies School of Health Research, Darwin, Northern Territory, Australia..
Global & Tropical Health Division, Menzies School of Health Research, Darwin, Northern Territory, Australia.. Faculty of Science, Health, Education & Engineering, University of the Sunshine Coast, Sippy Downs, Queensland, Australia..
Department of Molecular Genetics & Microbiology, College of Medicine, Emerging Pathogens Institute, University of Florida, Gainesville, FL, USA..
Department of Molecular Genetics & Microbiology, College of Medicine, Emerging Pathogens Institute, University of Florida, Gainesville, FL, USA..
Departments of Infectious Diseases & Pathology & Northern Territory Medical Program, Royal Darwin Hospital, Darwin, Northern Territory, Australia..
Global & Tropical Health Division, Menzies School of Health Research, Darwin, Northern Territory, Australia.. Departments of Infectious Diseases & Pathology & Northern Territory Medical Program, Royal Darwin Hospital, Darwin, Northern Territory, Australia..
Global & Tropical Health Division, Menzies School of Health Research, Darwin, Northern Territory, Australia.. Faculty of Science, Health, Education & Engineering, University of the Sunshine Coast, Sippy Downs, Queensland, Australia..
Issue Date: Sep-2018
Citation: Future microbiology 2018-09; 13: 1403-1418
Abstract: 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. 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. The triplex assay accurately detected efflux pump upregulation in all clinical and laboratory mutants, which corresponded with decreased antibiotic susceptibility or antibiotic resistance. 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.
URI: http://docs.prosentient.com.au/prosentientjspui/handle/1/11281
DOI: 10.2217/fmb-2018-0155
Type: Journal Article
Subjects: Burkholderia pseudomallei
antibiotic resistance
efflux pump
gene expression
melioidosis
qPCR
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