The increase of Beta-lactamases producing organisms can cause major therapeutic failure and poses a significant clinical challenge in healthcare settings. A total of 185 clinical isolates of Pseudomonas aeruginosa strains were collected from in-and out-patients at The Nairobi Hospital, 74.1 % were inpatients and 25.9% were outpatients with the high prevalence of this bacterium among the male gender (61.1%) than female(38.9%); and preponderantly comprising the patients above 45 years old (64.3%). The highest numbers of P. aeruginosa were isolated from pus swab (39.5%), respiratory secretions (25.9%), and urine (18.9%). The resistance rate of P. aeruginosa against carbapenem was 31.5% among the isolates. The prevalence of MBL producing P. aeruginosa was 22.7% as compared to non-MBL isolates (77.3%). The MBL isolates were resistant to the examined antibiotics. There were two predominant genes VIM-2 (28.57%) and NDM-1 (66.67%) types among MBL P. aeruginosa, and more prevalent genes were isolated from Critical care nursing ward; Intensive Care Unit (45.2%) and High Dependency Unite (28.6%) at The Nairobi Hospital. These findings suggest that the early detection of Metallo-Beta-Lactamases-producing isolates and the cooperation between medical professionals and infection control team may help in appropriate antimicrobial therapy and avoid further spread of these multidrug resistance strains. Keywords: Pseudomonas aeruginosa, Metallo-Beta-Lactamases, Resistance, Beta-lactams
