OBJECTIVES: Increasing colonization by beta-lactam-resistant Gram-negative bacilli (BR-GNB) represents a risk for infections and bacterial resistance spread, both in hospitals and the community. Hemodialysis patients and their household contacts regularly transit between these environments. This study investigated the clinical and epidemiological characteristics of BR-GNB colonization in hemodialysis patients and their household contacts, as well as the genetic relationship between their isolates. METHODS: A cross-sectional study was conducted on hemodialysis patients at a hospital-associated dialysis center in Medellín, Colombia and their household contacts. Clinical and epidemiological information was collected. Colonization was assessed from stool or rectal swab samples. Bacterial identification and susceptibility were determined using chromogenic media and Vitek-2. Molecular characterization included beta-lactamase detection by polymerase chain reaction, multiple-locus sequence typing (MLST), pulsed-field gel electrophoresis, and identification of Escherichia coli phylogroups by the Clermont protocol. RESULTS: This study included 36 hemodialysis patients and 90 household contacts. Colonization by BR-GNB occurred in 58.3% of patients and 22.2% of household contacts. The main beta-lactamase detected was CTX-M group-1 (40.5%). In 3 of the 9 homes that had more than 1 colonized individual, a genetic relationship was found. MLST showed a high diversity in E. coli isolates, and the most frequent phylogroups were B1 and B2. CONCLUSIONS: These results show a high frequency of colonization and the presence of potentially pathogenic BR-GBN both in hospitals and the community. This highlights the importance of populations who move between those 2 environments, and the need to prevent the spread of bacterial resistance outside hospitals.