Escherichia coli Ol 57:H7 serotype is worldwide zoonotic pathogenresponsible for the majority of severe cases of human enterohemorrhagic Escherichia coli. A cross sectional study was designed from November 2018-May 2019 to isolate, molecularly characterize E. coli serotype 0157:H7 and to assess its antimicrobial resistance pattern from rmv beef ready to eat "kurt "from butcher shop at Bahir Dar City. A total of 7 4 beef samples were collected and processed using standard microbiological procedures to isolate and identify the bacterium and determine their antimicrobial susceptibility patterns. Genomic DNA extracted from the isolates were examined in Polymerase Chain Reactions (PCR) to detect virulence genes (stxi and hlyA). Accordingly 21 E. coli Ol 57:H7 were isolated using compact dryEC kit and sorbitol maconky agar of which 61.9% stx, and 38.1%hlyA. Isolates were further subjected to the invitro antimicrobial sensitivity test and resulted in a degree of susceptibility ranged from 62% - 100% and resistance were between 4.8% and 33.3%. Only 6 (28.6%) of them showed the presence of resistance for two drugs amoxicillin and chloramphenicol. Finally the higher incidence of the stx, and hlyA gene isolated from meat samples in the butcher shop indicates a risk alert of E. coli 0157 infection. Therefore, appropriate measures are now needed to wards food safety.knowledge gap of butcher shop workers and building standardized abattoir in the area to prevent the spread of this life-threatening food borne disease in our city and our country Ethiopia having the habit eating uncooked beef