Introduction: Pseudomonas aeruginosa is a Gram-negative rod and an opportunistic pathogen that causes infections in hospitalized patients, some of them with an special issue to treat due to their ability to become resistant to almost all the families of antibiotics available. One of its main mechanisms of resistance to carbapenems is the acquisition and production of carbapenemases, enzymes capable of hydrolyzing most β-lactam antibiotics. KPC enzyme, class A carbapenemase most frequently found in Enterobacteriaceae and widely distributed in Klebsiella pneumoniae. However, in Colombia, P. aeruginosa isolates were detected for the first time with this enzyme and since then, it has spread throughout the country. The objective of this study was to determine the genetic diversity of P. aeruginosa isolates that cause infection in patients treated in tertiary level hospitals in Bogotá, Colombia. Methodology: A descriptive observational cross-sectional study was carried out in adult patients treated at four third-level institutions in the city of Bogotá. The genetic relationship was determined by PFGE and complete genome sequencing of two isolates was established by PacBio. Results: During the study period, 23 isolates of P. aeruginosa resistant to carbapenems and carriers of blaKPC gene were identified. Of these, 19 (83%) had blaKPC-2 variant; and 2 blaKPC-3; blaVIM gene was detected in 3 (12%). 100% of isolates were multiresistant. PFGE analysis revealed 12 pulsotypes distributed in 8 different clones. Clone 1 was the most frequent and identified in the 4 institutions, suggesting a wide spread. Two sequenced isolates belonged to ST111 and ST235 (pandemic clones) and showed two new platforms for mobilization of blaKPC gene related to Tn3 transposon and ISPae38 insertion sequence. For the first time, the mobilization and acquisition of blaKPC-3 gene is detected in P. aeruginosa worldwide. Conclusion: Pseudomonas aeruginosa has acquired, maintained and assimilated carbapenemase KPC. The behavior of the enzyme activity in this pathogen, as its clinical implications and activity to new anti-infective agents are unknown, constituting a new challenge due to the impact on molecular diagnosis of resistance and for the clinician, due to its results in manifestations, prognosis and treatment. Key words: Pseudomonas aeruginosa, blaKPC-3, Carbapenems, Resistance, genetic variability, Colombia