Purpose: To investigate the frequency of MBLs in Pseudomonas and Acinetobacter species with E-test, to determine the risk factors and to evaluate the demographic and clinical features of infected patients. Materials and methods: Imipenem or meropenem resistance of Pseudomonas and Acinetobacter isolated from several clinical samples with conventional methods were evaluated with imipenem EDTA E-test and the presence of Metallo-β-lactameses MBL was examined. Several isolates were screened for VIM-1, VIM-2, IMP-1, and IMP-2 with a PCR test. Results: Of 46 carbapenem resistant Acinetobacter isolates, 41 (89%), as well as of 19 carbapenem resistant Pseudomonas isolates, 5 (26%) had MBL positivity with imipenem-EDTA E-test. A history of Intensive Care Unit stay, mechanical ventilation and cephalosporin use were found to be significant risk factors with respect to MBL production. Conclusion: Detection of MBL production in Acinetobacter and Pseudomonas species especially in ICU patients is of prime importance to control infection rapidly and effectively, which contribute to prevention of outbreaks.