Background: Catheter -associated urinary tract infections (CAUTIs) are prevalent among intensive care unit (ICU) patients and pose significant health risks. The causative agent, Escherichia coil, is commonly associated with these infections. Objectives: The primary objective of our study was to evaluate various risk factors associated with CAUTIs. Additionally, we investigated the capacity of E. toil to form biofilms and the presence of virulence genes in these infections. Methods: We conducted a prospective study involving 204 patients admitted to ICUs with indwelling urinary catheters who subsequently developed CAUTIs. The isolated pathogen in all cases was E. coll. We subjected the isolated E. call strains to microbiological analyses, including biofilm formation assays and polymerase chain reaction (PCR) for detecting virulence genes (pap -C, fim-H, sfa, afa, hylA, and cnfi). Results: Our findings revealed that 111 out of 204 isolated E. call strains (54.4%) could form biofilms in vitro. Among the detected virulence genes, fim-H was the most frequently observed (32.4%), followed by pap -C (14.2%), sfa (123%), and afar (113%). The least frequently detected genes were cnft (3.9%) and hylA (2.5%). Notably, there was a significant association between the presence of fim-H, pap -C, afa1, and hylA genes and the ability of E. coil to form biofilms (P = 0.001, P = 0.001, P = 0.04, P = 0.046, respectively). Regarding the timing of CAUTIs, most cases occurred 7 -10 days after catheter insertion (48.53%), followed by 3 - 6 days after insertion (4559%). The least frequent occurrence was after 10 days from catheter insertion (5.86%). Early presence of CAUTIs (within 3 - 6 days) was significantly associated with diabetes mellitus (DM) and post -operative complications (both with P = 0.046). Furthermore, we observed a significant increase in the presence of the cnfi gene in cases of early CAUTIs (3 - 6 days after catheter insertion, P= 0.001). Conclusions: Our study highlights the importance of understanding the risk factors, virulence genes, and biofilm formation capacity in E. coil -related CAUTIs. These findings may guide preventive strategies and improve patient outcomes.