Files and links (2)
Rapid fluoroquinolone resistance detection in Pseudomonas aeruginosa using mismatch amplification mutation assay-based real-time PCR314.05 kB
Preprint VersionCC BY-NC-ND V4.0, Open Access
Preprint
Rapid fluoroquinolone resistance detection in Pseudomonas aeruginosa using mismatch amplification mutation assay-based real-time PCR
medRxiv, Vol.1 January 2022
Cold Spring Harbor Laboratory Press
2022
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 clinical P. aeruginosa isolates from Queensland, Australia, 45 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 (84%) of isolates with intermediate/full FQ resistance. Importantly, 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 Madden (Author) - University of the Sunshine Coast, Queensland, GeneCology Research Centre - LegacyKate L McCarthy (Author) - The University of QueenslandScott C Bell (Author) - The University of QueenslandOlusola Olagoke (Author) - University of the Sunshine Coast, Queensland, GeneCology Research Centre - LegacyTimothy Baird (Author) - Sunshine Coast University Hospital (Australia)Jane Neill (Author) - Sunshine Coast University Hospital (Australia)Kay A Ramsay (Author) - The University of QueenslandTimothy J Kidd (Author) - The University of QueenslandAdam G Stewart (Author) - The University of QueenslandShradha Subedi (Author) - Sunshine Coast University Hospital (Australia)Keat Choong (Author) - Sunshine Coast University Hospital (Australia)Tamieka Fraser (Author) - University of the Sunshine Coast, Queensland, School of Science and Engineering - LegacyDerek Sarovich (Author) - University of the Sunshine Coast, Queensland, GeneCology Research Centre - LegacyErin Price (Author) - University of the Sunshine Coast, Queensland, GeneCology Research Centre - Legacy
- Publication details
- medRxiv, Vol.1 January 2022
- Publisher
- Cold Spring Harbor Laboratory Press
- Date published
- 2022
- DOI
- 10.1101/2021.12.12.21266792
- Copyright note
- The copyright holder for this preprint is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC 4.0 International license.
- Organisation Unit
- School of Science and Engineering - Legacy; University of the Sunshine Coast, Queensland; School of Science, Technology and Engineering; Centre for Bioinnovation; School of Health and Behavioural Sciences - Legacy
- Language
- English
- Record Identifier
- 99627440702621
- Output Type
- Preprint
Metrics
96 Record Views