Background: Carbapenemases production by extended-spectrum β-lactamases (ESβLs)-producing Escherichia coli (E. coli) has been increasingly found and may be considered as a major cause of morbidity and mortality in hospital-acquired infection. Objectives: To determine resistance pattern and frequency of carbapenem resistance and presence of class D carbapenemases among ESβLs-producing E. coli isolated from patients in Menoufia University Hospitals. Methodology: Different clinical samples were obtained from 270 patients who were admitted to Menoufia University Hospitals. E. coli were isolated and identified, and their antimicrobial resistance profiles were tested by the disk diffusion and agar dilution methods. Confirmed ESβLs producers (by cephalosporin/ clavulanate combination disks and ESβL NDP tests) were further tested for carbapenemase production by phenotypic and genotypic methods. Results: E. coli was the most common isolate (30.4%) from clinical samples. High rate of ESβLs-producing E. coli was detected by disk diffusion (87.5%), cephalosporin/clavulanate combination disk (62.5%) and ESβL NDP test (60%).About10%, 24% and 28% of the ESβLs-producing E. coli isolates were producers of class A, B and D carbapenemases respectively. The prevalence of blashv, blaoxa23 and blaoxa48 genes among ESβLs-producing E. coli isolates was 18%, 22% and 12% respectively. Conclusion: Carbapenemases production by EβSLs-producing E. coli is a major challenge. A great concern should be paid to provide alternative new therapeutic agents, continuous surveillance, and effective antibiotic stewardship program.