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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.
- Received:
- Accepted:
- Published Online:
Funding
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University of the Sunshine Coast
- Principle Award Recipient: MaddenDanielle E.
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Wishlist
(Award 2019-14)
- Principle Award Recipient: FraserTamieka A.
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Wishlist
(Award 2019-14)
- Principle Award Recipient: PriceErin P.
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Wishlist
(Award 2019-14)
- Principle Award Recipient: SarovichDerek S.
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Wishlist
(Award 2019-14)
- Principle Award Recipient: BairdTimothy
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Advance Queensland
(Award AQRF13016-17RD2)
- Principle Award Recipient: SarovichDerek S.
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Advance Queensland
(Award AQIRF0362018)
- Principle Award Recipient: PriceErin P.