Jessica R Webb1, Erin P Price1,2, Nawarat Somprasong3, Herbert P Schweizer3, Robert W Baird4, Bart J Currie1,4, Derek S Sarovich1,2. 1. Global & Tropical Health Division, Menzies School of Health Research, Darwin, Northern Territory, Australia. 2. Faculty of Science, Health, Education & Engineering, University of the Sunshine Coast, Sippy Downs, Queensland, Australia. 3. Department of Molecular Genetics & Microbiology, College of Medicine, Emerging Pathogens Institute, University of Florida, Gainesville, FL, USA. 4. Departments of Infectious Diseases & Pathology & Northern Territory Medical Program, Royal Darwin Hospital, Darwin, Northern Territory, Australia.
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.
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.
Authors: Derek S Sarovich; Jessica R Webb; Matthew C Pitman; Linda T Viberg; Mark Mayo; Robert W Baird; Jennifer M Robson; Bart J Currie; Erin P Price Journal: Clin Infect Dis Date: 2018-07-02 Impact factor: 9.079
Authors: Nicole L Podnecky; Katherine A Rhodes; Takehiko Mima; Heather R Drew; Sunisa Chirakul; Vanaporn Wuthiekanun; James M Schupp; Derek S Sarovich; Bart J Currie; Paul Keim; Herbert P Schweizer Journal: mBio Date: 2017-09-05 Impact factor: 7.867
Authors: Derek S Sarovich; Erin P Price; Danielle E Madden; Olusola Olagoke; Timothy Baird; Jane Neill; Kay A Ramsay; Tamieka A Fraser; Scott C Bell Journal: Antimicrob Agents Chemother Date: 2022-04-25 Impact factor: 5.938