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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,\n \n \n Pseudomonas aeruginosa<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic>\n .\n <\/jats:p>\n \n Objectives.<\/jats:bold>\n We aimed to develop two SYBR Green-based mismatch amplification mutation assays (SYBR-MAMAs) targeting GyrA T83I (\n gyrA<\/jats:italic>\n 248) and GyrA D87N, D87Y and D87H (\n gyrA<\/jats:italic>\n 259). Together, these variants cause the majority of fluoroquinolone (FQ) AMR in\n \n \n P. aeruginosa<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic>\n .\n <\/jats:p>\n \n Methods.<\/jats:bold>\n Following assay validation, the\n gyrA<\/jats:italic>\n 248 and\n gyrA<\/jats:italic>\n 259 SYBR-MAMAs were tested on 84 Australian clinical\n \n \n P. aeruginosa<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic>\n isolates, 46 of which demonstrated intermediate\/full ciprofloxacin resistance according to antimicrobial susceptibility testing.\n <\/jats:p>\n \n Results.<\/jats:bold>\n 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\u200a% concordance with SYBR-MAMA genotypes.\n <\/jats:p>\n \n Conclusions.<\/jats:bold>\n Our GyrA SYBR-MAMAs provide a rapid and cost-effective method for same-day identification of FQ AMR in\n \n \n P. aeruginosa<\/jats:ext-link>\n <\/jats:named-content>\n <\/jats:italic>\n . An additional SYBR-MAMA targeting the GyrB S466Y\/S466F variants would increase FQ AMR prediction to 91\u200a%. 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.\n <\/jats:p>","DOI":"10.1099\/jmm.0.001593","type":"journal-article","created":{"date-parts":[[2022,10,27]],"date-time":"2022-10-27T12:19:55Z","timestamp":1666873195000},"source":"Crossref","is-referenced-by-count":8,"title":["Rapid fluoroquinolone resistance detection in Pseudomonas aeruginosa using mismatch amplification mutation assay-based real-time PCR"],"prefix":"10.1099","volume":"71","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3261-750X","authenticated-orcid":false,"given":"Danielle E.","family":"Madden","sequence":"first","affiliation":[{"name":"Centre for Bioinnovation, University of the Sunshine Coast, Sippy Downs, Queensland, Australia"},{"name":"Infection Research Network Sunshine Coast, Birtinya, Queensland, Australia"},{"name":"Sunshine Coast Health 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