This study was conducted to obtain data about prevalence of plasmid-mediated AmpC (pAmpC), efficacy of various phenotypic tests applied for the detection of pAmpC-mediated resistance and the enzyme types responsible for this resistance. A total of 1316 isolates (1030 Escherichia coli and 286 Klebsiella spp.) obtained from the clinical samples sent to our laboratory between 2008-2011 period, from various inpatient and outpatient clinics and intensive care units, were included in this study. Antimicrobial susceptibility profiles of the isolates were determined by using Kirby-Bauer disk diffusion method. Production of pAmpC was phenotypically investigated by Boronic Acid Inhibition Test (BAIT), Cefoxitin Hodge Test (CHT) and AmpC Disk Test (ACDT) in a total of 36 (2.7%) cefoxitin-resistant isolates (77% were E.coli, 23% were K.pneumoniae); and by synergy tests with or without AmpC inhibitors in a total of 165 (88% were E.coli, 12% were K.pneumoniae) cefoxitin-susceptible, third generation cephalosporin-resistant (S3R) isolates. For the detection of pAmpC genes a multiplex-PCR protocol was applied to the isolates found positive at least by one of the phenotypic tests. CHT, ACDT and BAIT were found positive in 21 (58%), 18 (50%) and 7 (19%) of the 36 cefoxitin-resistant isolates, respectively. In only 10 (27.7%) of these isolates (all were E.coli strains), pAmpC presence was detected by PCR; and the enzyme produced was assessed as CMY-2. Based on the positive PCR results; specificity rates of the phenotypic tests, BAIT, ACDT and CHT were 97%, 69% and 58%; while the sensitivity rates were 50%, 100% and 100%, respectively. Our data indicated that, pAmpC prevalence (10/1316, 0.8%) detected in this study, did not seem to be an important problem yet, in the population screened. However, since the first detection of this resistance was in 2010, it should be considered as a sign to get necessary precautions preventing its spread. In conclusion, none of the phenotypic methods were satisfactory alone for the detection of pAmpC-mediated resistance; and BAIT was superior in terms of specificity while the others were superior in terms of sensitivity. Thus, combined application of these phenotypic tests are necessary to screen and confirm the presence of pAmpC-mediated resistance.