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Published VersionCC BY V4.0, Open Access
Journal article
Rapid fluoroquinolone resistance detection in Pseudomonas aeruginosa using mismatch amplification mutation assay-based real-time PCR
Journal of Medical Microbiology, Vol.71, pp.1-10
2022
Appears in UniSC Supported Open Access Outputs
Abstract
Background. Antimicrobial resistance (AMR) is an ever-increasing global health concern. One crucial facet in tackling the AMR epidemic is earlier and more accurate AMR diagnosis, particularly in the dangerous and highly multi-drug-resistant ESKAPE pathogen, Pseudomonas aeruginosa.
Objectives. We aimed to develop two SYBR Green-based mismatch amplification mutation assays (SYBR-MAMAs) targeting GyrA T83I (gyrA248) and GyrA D87N, D87Y and D87H (gyrA259). Together, these variants cause the majority of fluoroquinolone (FQ) AMR in P. aeruginosa.
Methods. Following assay validation, the gyrA248 and gyrA259 SYBR-MAMAs were tested on 84 Australian clinical P. aeruginosa isolates, 46 of which demonstrated intermediate/full ciprofloxacin resistance according to antimicrobial susceptibility testing.
Results. Our two SYBR-MAMAs correctly predicted an AMR phenotype in the majority (83%) of isolates with intermediate/full FQ resistance. All FQ-sensitive strains were predicted to have a sensitive phenotype. Whole-genome sequencing confirmed 100 % concordance with SYBR-MAMA genotypes.
Conclusions. Our GyrA SYBR-MAMAs provide a rapid and cost-effective method for same-day identification of FQ AMR in P. aeruginosa. An additional SYBR-MAMA targeting the GyrB S466Y/S466F variants would increase FQ AMR prediction to 91 %. Clinical implementation of our assays will permit more timely treatment alterations in cases where decreased FQ susceptibility is identified, leading to improved patient outcomes and antimicrobial stewardship.
Details
- Title
- Rapid fluoroquinolone resistance detection in Pseudomonas aeruginosa using mismatch amplification mutation assay-based real-time PCR
- Authors
- Danielle E. Madden (Author) - University of the Sunshine Coast, Queensland, Centre for BioinnovationKate L. McCarthy (Author) - The University of QueenslandScott C. Bell (Author) - Prince Charles HospitalOlusola Olagoke (Author) - University of the Sunshine Coast, Queensland, Centre for BioinnovationTimothy Baird (Author) - University of the Sunshine Coast, Queensland, Centre for BioinnovationJane Neill (Author) - Sunshine Coast University HospitalKay A. Ramsay (Author) - The University of QueenslandTimothy J. Kidd (Author) - Royal Brisbane and Women's HospitalAdam G. Stewart (Author) - Sunshine Coast University HospitalShradha Subedi (Author) - Sunshine Coast University HospitalKeat Choong (Author) - Sunshine Coast University HospitalTamieka A. Fraser (Author) - University of the Sunshine Coast, Queensland, Centre for BioinnovationDerek S. Sarovich (Author) - University of the Sunshine Coast, Queensland, Centre for BioinnovationErin P. Price (Corresponding Author) - University of the Sunshine Coast, Queensland, Centre for Bioinnovation
- Publication details
- Journal of Medical Microbiology, Vol.71, pp.1-10
- Publisher
- The Microbiology Society
- Date published
- 2022
- DOI
- 10.1099/jmm.0.001593
- ISSN
- 1473-5644; 0022-2615
- Copyright note
- © 2022 The Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution NonCommercial License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
- Grant note
- This work was supported by Advance Queensland (awards AQIRF0362018 and AQRF13016-17RD2), an Australian Government Research Training Program Scholarship, the Wishlist Sunshine Coast Health Foundation (award 2019-14), and the National Health and Medical Research Council (award 455919). The funders had no role in study design, data acquisition, analysis, interpretation, writing or submission of the manuscript.
- Organisation Unit
- School of Science and Engineering - Legacy; School of Science, Technology and Engineering; Centre for Bioinnovation; School of Health and Behavioural Sciences - Legacy
- Language
- English
- Record Identifier
- 99716027502621
- Output Type
- Journal article
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