Hospital wastewater is believed to be a leading source of antibiotic resistant bacteria, which presents a risk to both the ecosystem and public health. The current study endeavored to isolate and identify Escherichia coli from the wastewater of multiple hospitals in Baghdad City, Iraq, and characterize its antibiotic resistance patterns. The study resulted in identifying, by biochemical means using the VITEK system, a total of 50 E.coli isolates. Antibiotic susceptibility tests for 11 antibiotics were conducted on each isolate via the Kirby–Bauer (KB) disk diffusion method. Overall, all isolates showed considerable resistance to the following antibiotics: erythromycin and clarithromycin (100%), amoxicillin (86%), cefoxitin (72%), ceftriaxone (66%), and tetracycline (56%). Conversely, gentamicin (6%) and meropenem (16%) exhibited the lowest resistance patterns among all isolates. The Multiple Antibiotic Resistance (MAR) index indicated that 44 isolates (88%) exceeded 0.2, signifying that a high number of isolates were contaminated or exposed to antibiotics