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

2 Downloads (Pure)

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
Pages (from-to)	1403-1418
Number of pages	16
Journal	Future Microbiology
Volume	13
Issue number	12
Early online date	26 Sep 2018
DOIs	https://doi.org/10.2217/fmb-2018-0155
Publication status	Published - Sep 2018

Access to Document

10.2217/fmb-2018-0155

21246627-oaSubmitted manuscript, 4.06 MBLicence: CC BY-NC-ND

Cite this

@article{77ddb28ca4d242fdb8819f4983227ca2,

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 = "antibiotic resistance, Burkholderia pseudomallei, 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.}",

year = "2018",

month = sep,

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

language = "English",

volume = "13",

pages = "1403--1418",

journal = "Future Microbiology",

issn = "1746-0913",

publisher = "Global Science Books",

number = "12",

}

Development and validation of a triplex quantitative real-time PCR assay to detect efflux pump-mediated antibiotic resistance in Burkholderia pseudomallei. / Webb, Jessica R.; Price, Erin P.; Somprasong, Nawarat; Schweizer, Herbert P.; Baird, Robert W.; Currie, Bart J.; Sarovich, Derek S.

In: Future Microbiology, Vol. 13, No. 12, 09.2018, p. 1403-1418.

Research output: Contribution to journal › Article › peer-review

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 - antibiotic resistance

KW - Burkholderia pseudomallei

KW - efflux pump

KW - gene expression

KW - melioidosis

KW - qPCR

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

U2 - 10.2217/fmb-2018-0155

DO - 10.2217/fmb-2018-0155

M3 - Article

C2 - 30256166

AN - SCOPUS:85054448678

VL - 13

SP - 1403

EP - 1418

JO - Future Microbiology

JF - Future Microbiology

SN - 1746-0913

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

Access to Document

Other files and links

Fingerprint

Cite this