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

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 journal › Article › peer-review

7 Scopus citations

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 language	English (US)
Pages (from-to)	1403-1418
Number of pages	16
Journal	Future Microbiology
Volume	13
Issue number	12
DOIs	https://doi.org/10.2217/fmb-2018-0155
State	Published - Sep 2018
Externally published	Yes

Keywords

Burkholderia pseudomallei
antibiotic resistance
efflux pump
gene expression
melioidosis
qPCR

ASJC Scopus subject areas

Microbiology
Microbiology (medical)

Access to Document

10.2217/fmb-2018-0155

Cite this

@article{1b2892e329fc4125a92f513f5fa76547,

title = "Development and validation of a triplex quantitative real-time PCR assay to detect efflux pump-mediated antibiotic resistance in Burkholderia pseudomallei",

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.",

keywords = "Burkholderia pseudomallei, antibiotic resistance, efflux pump, gene expression, melioidosis, qPCR",

author = "Webb, {Jessica R.} and Price, {Erin P.} and Nawarat Somprasong and Schweizer, {Herbert P.} and Baird, {Robert W.} and Currie, {Bart J.} and Sarovich, {Derek S.}",

note = "Publisher Copyright: {\textcopyright} 2018 2018 Future Medicine Ltd.",

year = "2018",

month = sep,

doi = "10.2217/fmb-2018-0155",

language = "English (US)",

volume = "13",

pages = "1403--1418",

journal = "Future Microbiology",

issn = "1746-0913",

publisher = "Future Medicine Ltd.",

number = "12",

}

TY - JOUR

T1 - Development and validation of a triplex quantitative real-time PCR assay to detect efflux pump-mediated antibiotic resistance in Burkholderia pseudomallei

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.

PY - 2018/9

Y1 - 2018/9

N2 - 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.

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.

KW - Burkholderia pseudomallei

KW - antibiotic resistance

KW - efflux pump

KW - gene expression

KW - melioidosis

KW - qPCR

UR - http://www.scopus.com/inward/record.url?scp=85054448678&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85054448678&partnerID=8YFLogxK

U2 - 10.2217/fmb-2018-0155

DO - 10.2217/fmb-2018-0155

M3 - Article

C2 - 30256166

AN - SCOPUS:85054448678

SN - 1746-0913

VL - 13

SP - 1403

EP - 1418

JO - Future Microbiology

JF - Future Microbiology

IS - 12

ER -

Development and validation of a triplex quantitative real-time PCR assay to detect efflux pump-mediated antibiotic resistance in Burkholderia pseudomallei

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this