A total of 106 gram negative bacteria isolates obtained from culture of various specimens of patients hospitalized in YGH in March and April 2015 were studied. Sixty-two gram-negative bacteria isolates resistant to broad spectrum cephalosporins consisting of 51 Enterobacteriaceae and 11 nonfermenting β lactamase producing gram-negative bacteria isolates were confirmed by conventional methods, RapID Systems for Enterobacteriaceae and nonfermenting bacteria and fluorescence in situ hybridization (FISH) for one isolate of nonfermenting E. coli and Acinetobacter species were carried out. Antimicrobial susceptibility pattern were confirmed with disk diffusion method, according to the CLSI guidelines. The ESBL phenotype was confirmed on a culture plate, using the disk approximation test. Extensively drug-resistant (XDR) isolates were 10.5% (n=4) of Escherichia coli and 30% (n=3) of Klebsiella pneumoniae. ESBL-producing isolates represented 68% (n=26) of E. coli and 60% (n=6) of K. pneumoniae. A Pseudomonas aeruginosa isolate was found to be pan drug-resistant (PDR) to all tested antimicrobials. All Acinetobacter species isolates (n=4, 100%) were found to be resistant to ampicillin, aztreonam, cephalothin, cefotaxime, cloramphenicol, cefoxitin, and streptomycin. All P. aeruginosa isolates were found to be resistant to tobramycin, ciprofloxacin, levofloxacin, ceftazidime, cefepime, meropenem, and ticarcillin and (83%) were resistant to aztreonam and piperacillintazobactam, three (50%) were resistant to imipenem and piperacillin. Imipenem resistance was 10.5% (n=4) in Escherichia coli, 30% (n=3) in Klebsiella pneumoniae, 33.3% (n=1) in Enterobacter cloacae, 50% (n=2) in Acinetobacter spp., 50% (n=3) in Pseudomonas aeruginosa.