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Abstract The urinary tract infection is one of the most common infectious diseases. The increasing prevalence of Multi-drug resistant (MDR) organisms worldwide results inthe availability of limitedoptions for the treatment of infections caused by such resistant pathogens. Recently, physicians have reconsidered the use of old antibiotics to overcome this problem. Fosfomycin is one of these antibiotics and has showed a good in vitro activity against extended spectrum Ý-lactamase (ESBL)-producing bacteria and multidrug-resistant(MDR) organisms in many studies. This study aims to assess the in vitro activity of fosfomycin against ESBL-producing MDR uropathogens. Over one year we collected 472 ESBL-producing MDR isolates from the urinary samples delivered to the microbiology laboratory of Cairo University hospitals. ESBL production was confirmed by the double disk synergy test, the Cephalosporin/Clavulanate combination disk test, and ESBL-chromogenic medium. Fosfomycin susceptibility testing was done using disk diffusion test. Among the 472 isolates, 187 (39.6%) were Klebsiella species, 146 (30.9%) were E. coli, 89 (18.9%) were Pseudomonas spp., 38 (8.1%) were Acinetobacter, and 12 (2.5%) isolates were others including Enterobacter, Morganella, Providencia, and Proteus spp. Fosfomycin was found to have 69.1% (326/472) sensitivity rate among all isolates with a significant higher sensitivity rate among E. coli 93.1% (136/146). Conclusion: Fosfomycin showed in vitro activity against 69.1% of the ESBL-producing MDR uropathogens. E. coli showed statistically significantly higher sensitivity rate to fosfomycin than other tested species |