Acinetobacter baumannii is an opportunistic pathogen causing infections in hospitalized patients, particularly in intensive care units (ICUs). The treatment of these infections is one of the most important concerns of physicians due to multidrug-resistant. The b-lactam antimicrobial drugs are choice treatment of infections caused by A. baumannii. The specimens included CSF (cerebrospinal fluid), wound, sputum, urine, BAL (Broncho Alveolar Lavage), blood and so on were taken from 128 patients who were hospitalized in several wards. The antimicrobial susceptibility test was carried out using kirby-bauer disc diffusion method for Piperacillin-Tazobactam, Ceftriaxon, Ceftazidime, Cefepime, Imipenem, Doripenem, Ertapenem, Cefotaxime, Ampicillin-Sulbactam antibiotics for all isolates. DNA of isolates was extracted and primers were designed and as well as amplification was carried out for blaIMP, blaVIM, blaNDM1 genes. The resistance to all discs used in this study was seen in all isolates. The Highest and the lowest rate of resistance were seen in Piperacillin-tazobactam and Ampicillin-sulbactam, 100% and 21% isolates, respectively. The results of PCR of blaIMP, blaVIM and blaNDM genes were as follows: blaIMP 5/128 (3.9%), blaVIM 9/128(7.03%) and blaNDM 0/128(0%). The highest sensitivity to antibiotics group was seen in Ampicillin-Sulbactam groups and also blaVIM was the most prevalent gene of resistance among Metallo-Beta-Lactamase genes in Acinetobacter baumannii isolates.