Phenotypic detection of plasmid-acquired AmpC in Escherichia coli--evaluation of screening criteria and performance of two commercial methods for the phenotypic confirmation of AmpC production.

The phenotypic detection of plasmid-acquired AmpC (pAmpC) in Escherichia coli is challenging, and molecular methods are required for confirmation. In addition to cefoxitin resistance, multiresistance and high-level resistance to cephalosporins have both been suggested as criteria for targeting isolates with pAmpC, but data to support these proposed criteria are lacking. A Swedish collection of 378 isolates with either putative chromosomal hyperproduction of AmpC (cAmpC) or pAmpC were subjected to disk diffusion and minimum inhibitory concentration (MIC) determination with the Etest. The frequency of resistance to gentamicin, ciprofloxacin, and trimethoprim among cAmpC and pAmpC was compared to elucidate the issue of multidrug resistance. Lastly, methods for the phenotypic confirmation of pAmpC were compared. One in-house disk diffusion method, one method employing NeoSensitabs (Rosco), and one Etest method (bioMérieux) were compared. The analysis of histograms based on both disk diffusion and the Etest showed that resistance [according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST)] to cefotaxime and/or ceftazidime occurred in almost all isolates. By coining resistance instead of non-susceptibility, the number of isolates required to subject to phenotypic testing/genotypic confirmation dropped by more than 40 %, without compromising the sensitivity substantially. Further, almost 70 % of isolates with pAmpC were non-multidrug resistant, clearly indicating that this is an inappropriate criterion for further investigation. The phenotypic tests all had more than 90 % sensitivity, and the best sensitivities were obtained with the in-house method and with the ceftazidime ± cloxacillin NeoSensitab. In conclusion, clinical resistance to cefotaxime and/or ceftazidime seems to be an appropriate criterion for pAmpC screening, and several phenotypic methods perform well for the phenotypic confirmation of AmpC production prior to genotypic confirmation.