Introduction: KPC enzymes are carbapenemases with a great capability to disseminate and to cause epidemics. They are frequently associated with higher mortality rates and prolonged hospital stay. In Colombia, they have been progressively reported since 2007; however, its prevalence in hospitals is not known.Objective: To estimate the prevalence of blaKPC gene in hospitals.Methods and materials: The presence of blaKPC gene and its clonality were evaluated in clinical isolates of Enterobacteriacea and Pseudomonas aeruginosa in hospitalized patients.Results: Of the 424 isolates tested during the study period, 273 met eligibility criteria, and 31.1% were positive for blaKPC gene; after clonality adjustment, positivity was 12.8%. The blaKPC gene was more frequent in Klebsiella pneumonia, followed by P. aeruginosa and other Enterobacteriacea. Although intensive care units (ICU) provided the majority of the isolates, the blaKPC pattern was not more prevalent in ICUs than in other wards. The respiratory tract was the anatomic source with the highest prevalence. No seasonality was observed associated with the frequency of isolation of microorganisms carrying blaKPC gene.Conclusion: This study revealed a high prevalence of blaKPC gene in microorganisms isolated from different hospitals in Colombia. The extraordinary ability of blaKPC gene to spread, the difficulties for its diagnosis and the limited antibiotics available for its treatment pose the urgent need to strengthen epidemiological surveillance systems, and to timely adjust institutional policies for rational use of antibiotics in order to limit its dissemination to other institutions in the country.