Abstract Escherichia coli in immunocompetent hosts is the causative agent of uncomplicated urinary tract infections and opportunistic infections in immunocompromised hosts and causes urinary infections. The minimum inhibitory MIC concentration among UPEC isolates ranged from 128 μg/ml to 512 μg/ml under this study. Molecular pattern of glpT and uhpT resistance factors genes toward fosfomycin in E. coli isolates of Iraqi UTI patients were determined in this review to clarify their role in fosfomycin uptake resistance system. Methods From the hospitals in Baghdad-Iraq, 50 urinary collected samples were obtained from fifty patients. The diagnosis was made using housekeeping gene 16sDNA gene. Minimum fosfomycin inhibitory concentrations (MICs) were tested the method of agar dilution with a concentrations between 16 and 1024 μg/mL. In addition, the investigation of glpT and uhpT genes was determined with specific primers using polymerase chain reaction PCR. Results The genetic identifying of Escherichia coli isolates with 16sDNA recorded all isolates gave positive results with amplified size of 544 bp for 16sDNA were recorded as specific gene detection for all isolates. In comparison to the break point, the highest MIC value was recorded about 256 μg/mL, 512 μg/mL and 1024 μg/mL which appeared resistance in fosfomycin ≤128 μg/ml. Moreover, the results showed that the presence of glpT and uhpT in all UPEC tested isolates which contributing in elevating the fosfomycin resistance by inducing the efflux pump system. Conclusion Our results showed highly susceptibility of UPEC isolates under this study toward the fosfomycin. We concluded, that the prevalence of glpT and uhpT It is considerably related to the progress of fosfomycin resistant by inducing efflux pump system to eliminate the drug out of the bacterial cells.