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Rapid fluoroquinolone resistance detection in Pseudomonas aeruginosa using mismatch amplification mutation assay-based real-time PCR

Antimicrobial resistance (AMR) is an ever-increasing global health concern. Here, we developed two SYBR Green-based mismatch amplification mutation assays (SYBR-MAMAs) targeting GyrA T83I, D87N, D87Y, and D87H, the predominant causes of fluoroquinolone AMR in Pseudomonas aeruginosa. SYBR-MAMAs were tested on 85 isolates, 45 with intermediate/full ciprofloxacin resistance. Assays identified the correct phenotype in 84% intermediate/full resistant and 100% sensitive strains. Clinical implementation of our rapid SYBR-MAMAs will permit timely treatment alterations to improve patient outcomes.