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Resistance to third generation cephalosporins is widely disseminated in Enterobacteriaceae mainly because of extended-spectrum-β-lactamases (ESBL), plasmid AmpC β-lactamases (PABL), and hyper-production of chromosomal AmpC β-lactamases. Here, we evaluated the performance of rapid test using novel fluorogenic probe assay in simulated blood cultures and compared the results with the phenol red assay using a total of 172 characterized isolates (39 ESBL producers, 13 PABL producers, and 120 susceptible isolates). We prepared a pellet by centrifugation and washing, which can also be used for identification with MALDI-TOF directly from positive blood cultures. After that, we mixed the pellet with fluorogenic probe and measured the fluorescent signal using fluorometer. The fluorogenic probe assay showed higher sensitivity than the phenol red assay (96.2% vs. 71.2%, p < .0001) in 172 simulated blood culture bottles especially in detecting PABL (84.6% vs. 0%, p = .0026) and the turnaround time was 1.5 h. This fluorogenic probe assay, combined with the direct identification of pathogens, could be very useful for rapid identification of isolates and detecting cephalosporin resistance caused by ESBL and PABL directly from positive blood cultures., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Resistance to third generation cephalosporins is widely disseminated in Enterobacteriaceae mainly due to extended-spectrum-β-lactamases, plasmid AmpC β-lactamases, and hyperproduction of chromosomal AmpC β-lactamases. Here we evaluated the performance of a novel fluorogenic probe rapid test and compared the results with the phenol red assay using a total of 77 characterized organisms (44 extended-spectrum-β-lactamases, 33 chromosomal or plasmid AmpC β-lactamases) and 46 susceptible organisms. The fluorescent assay showed higher sensitivity than the phenol red assay in cefotaximase type extended-spectrum-β-lactamases, non- cefotaximase type extended-spectrum-β-lactamases, chromosomal AmpC β-lactamases, and plasmid AmpC β-lactamases (96.7% vs. 90.0%, p=0.157; 71.4% vs. 7.1%, p=0.003; 100.0% vs. 64.7%, p<0.001; 100.0% vs. 6.3%, p<0.001). The fluorescent assay had a positive correlation with the exponents of cefotaxime and ceftazidime minimum inhibitory concentrations (p<0.001 for both). The new fluorescent assay will be very useful for the rapid detection of resistance to third generation cephalosporins that originates from various β-lactamases., (Copyright © 2016 Elsevier B.V. All rights reserved.)