Background:  Escherichia coli O157 is recognized as a major food-borne pathogen of great concern which can be transmitted by consumption of meat and meat products. Aim: This cross sectional study was conducted to determine the prevalence and antimicrobial resistance of E. coli O157 in mutton meat from butcher shops in Khartoum State, Sudan.  Methods: A total of 500 mutton meat carcass swabs were collected in the period March-December 2021 and processed according to ISO16654:2001 methods. Six antibiotics were used to evaluate antimicrobial susceptibility by disc diffusion method according to the guidelines of CLSI. Results: The results showed that the prevalence of E. coli O157 was 1% (5/500) from mutton meat. All isolates were sensitive to amikacin 5 (100%), while all isolates were resistant to ampicillin 5 (100%). For other antimicrobials, the resistance was cotrimoxazole 4 (80%), cefuroxime and cefotaxime 3 (60%). Three out of five isolates (60%) showed multiple drug resistance (MDR). Conclusions: The findings of this study demonstrated that E. coli O157 was present in mutton meat and frequently exhibited multiple drug resistance. Proper manufacturing processes and regular antimicrobial resistance monitoring should be taken into account,