Uropathogenic Escherichia coli (UPEC) is the main cause of urinary tract infections; in recent years, its importance as a pathogen has increased due to the emergence of hypervirulent and multiresistant strains. In this study, 190 urinary isolates of E. coli were assigned into the seven phylogenetic groups A (11.1%), B1 (4.7%), B2 (46.8%), C (5.8%) D (25.3%) F (2.6%), and Clade I (2.1%), and various virulence genes were examined with polymerase chain reaction methods. All isolates had at least one virulence factor of the 9 analyzed fyuA (81.1%), fimH (96.8%), iutA (74.7%), ompT (66.8%), kpsMTII (66.8%), traT (58.9%), PAI (43.6%), PapAH (26.3%), and usp (3.2%). The results showed a direct relationship between the virulence factors and phylogenetic group A and B2. Further, virulence genetic profiles fimH, fyuA, ompT, traT, and kpsMTII correlated with the production of strong biofilm, multidrug resistance, and the production of moderate hemolysin. These results suggest that these strains may become reservoirs of genes that encode virulence factors, which could be transferred horizontally enhancing their genomic background and high possibility of acquiring new genetic information for possible dissemination. This study provides the first description of phylogroups in UPEC in the Colombian Caribbean and the association with virulence factor profile, antimicrobial susceptibility, and their possible role in the epidemiology in Colombia.