Background/aim: This prospective study aimed to determine the presence of the most common carbapenemase genes, blaOXA-48, blaKPC, blaIMP, blaVIM and blaNDM on carbapenem resistant clinical K.pneumoniae and E.coli isolates. Materials and methods: Isolates were selected according to EUCAST guideline; gradient test and disc diffusion with both meropenem and ertapenem discs. Resistance rates of these isolates to other antimicrobial agents were also examined by disc diffusion method. Carbapenem resistance gene were investigated by using Real-Time PCR. Results: A total of 3845 E. coli and 1689 K. pneumoniae isolates from clinical samples between January 2015 and April 2017 were evaluated. The 419 isolates were found as carbapenem resistant but only the first resistant isolate (n=155; 126 K. pneumoniae and 29 E.coli) of each patient were included. Carbapenem resistant isolates were most frequently isolated from intensive care units (48.8%). Colistin was the most effective antibiotic (91.0%). The 121 (78.1%) of the tested isolates were positive for OXA-48 (103 K. pneumoniae and 18 E. coli) and 9 K. pneumoniae carrying blaNDM were also positive for blaOXA-48. VIM, IMP and KPC type carbapenemases were not detected in any isolates. Conclusion: Carbapenem-resistant pathogens have been shown to be able to develop resistance mechanisms with more than one carbapenemase encoding gene.