title: Rapid detection of AAC(6')-Ib-cr production using a MALDI-TOF MS strategy.
abstract: Plasmid-mediated quinolone resistance mechanisms have become increasingly prevalent among Enterobacteriaceae strains since the 1990s. Among these mechanisms, AAC(6')-Ib-cr is the most difficult to detect. Different detection methods have been developed, but they require expensive procedures such as Sanger sequencing, pyrosequencing, polymerase chain reaction (PCR) restriction, or the time-consuming phenotypic method of Wachino. In this study, we describe a simple matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) method which can be easily implemented in clinical laboratories that use the MALDI-TOF technique for bacterial identification. We tested 113 strains of Enterobacteriaceae, of which 64 harbored the aac(6')-Ib-cr gene. We compared two MALDI-TOF strategies, which differed by their norfloxacin concentration (0.03 vs. 0.5 g/L), and the method of Wachino with the PCR and sequencing strategy used as the reference. The MALDI-TOF strategy, performed with 0.03 g/L norfloxacin, and the method of Wachino yielded the same high performances (Se = 98 %, Sp = 100 %), but the turnaround time of the MALDI-TOF strategy was faster (<5 h), simpler, and inexpensive (<1 Euro). Our study shows that the MALDI-TOF strategy has the potential to become a major method for the detection of many different enzymatic resistance mechanisms.
authors: Laboratoire de Bactériologie Clinique, CHU Clermont-Ferrand, 58, rue Montalembert, 63003, Clermont-Ferrand, France.
authors: C-A
authors: CA
authors: Pardo
authors: Laboratoire de Bactériologie Clinique, CHU Clermont-Ferrand, 58, rue Montalembert, 63003, Clermont-Ferrand, France.
authors: R N
authors: RN
authors: Tan
authors: Laboratoire de Bactériologie Clinique, CHU Clermont-Ferrand, 58, rue Montalembert, 63003, Clermont-Ferrand, France. claire.hennequin@udamail.fr.|Centre National de Référence de la Résistance aux Antibiotiques, laboratoire associé, Clermont-Ferrand, France. claire.hennequin@udamail.fr.|LMGE-UMR CNRS 6023, Clermont Ferrand, 63000, France. claire.hennequin@udamail.fr.
authors: C
authors: C
authors: Hennequin
authors: Laboratoire de Bactériologie Clinique, CHU Clermont-Ferrand, 58, rue Montalembert, 63003, Clermont-Ferrand, France.|Centre National de Référence de la Résistance aux Antibiotiques, laboratoire associé, Clermont-Ferrand, France.
authors: R
authors: R
authors: Beyrouthy
authors: Laboratoire de Bactériologie Clinique, CHU Clermont-Ferrand, 58, rue Montalembert, 63003, Clermont-Ferrand, France.|Centre National de Référence de la Résistance aux Antibiotiques, laboratoire associé, Clermont-Ferrand, France.|Clermont Université, Université d'Auvergne, M2iSH, Clermont-Ferrand, France.|UMR INSERM 1071, Clermont-Ferrand, France.|USC INRA2018, Clermont-Ferrand, France.
authors: R
authors: R
authors: Bonnet
authors: Laboratoire de Bactériologie Clinique, CHU Clermont-Ferrand, 58, rue Montalembert, 63003, Clermont-Ferrand, France.|Centre National de Référence de la Résistance aux Antibiotiques, laboratoire associé, Clermont-Ferrand, France.|Clermont Université, Université d'Auvergne, M2iSH, Clermont-Ferrand, France.|UMR INSERM 1071, Clermont-Ferrand, France.|USC INRA2018, Clermont-Ferrand, France.
authors: F
authors: F
authors: Robin
mesh_terms: D000900:Anti-Bacterial Agents / Q000378:metabolism* / Q000494:pharmacology; D001431:Bacteriological Techniques / Q000379:methods*; D001711:Biotransformation; D024881:Drug Resistance, Bacterial; D004755:Enterobacteriaceae / Q000187:drug effects* / Q000378:metabolism; D009643:Norfloxacin / Q000378:metabolism* / Q000494:pharmacology; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013997:Time Factors
chemical_list: D000900:Anti-Bacterial Agents; D009643:Norfloxacin