E. coli O157 H7 is one of the most virulent foodborne pathogen. The aim of this study was to identify E coli O157 H7 from beef carcass samples at Bahir Dar municipal abattoir, determine antimicrobial resistance profiles of the isolates and to assess knowledge, attitude and practice of abattoir workers about the pathogen. A total of 280 swab samples were collected from carcass of cattle slaughtered at the abattoir and 25 (8.9%) isolates were identified by using sorbitol MacConkey agar supplemented with cefixime telluride and confirmed with latex agglutination test. A PCR was performed on isolates for the detection of virulence genes stx1, stx2, hlyA and eae genes. Antimicrobial susceptibility testing was performed using the disk diffusion method. Questionnaire was used to gather data about knowledge, attitude and practice of abattoir workers. Out of 25 isolates subjected for molecular detection 8 (32%) and 14 (56%) isolates possessed stx1 and stx2 genes respectively; from those 5(20%) isolates had both genes for the production of Shiga toxins. Relatively higher proportion; 18 (72%) isolates carried hlyA gene. Only 5(2%) isolates were positive for eae. Resistance was detected in All 25 (100%) isolates and 3(12%) against clindamycin and trimethoprim respectively. In addition, 3(12%) isolates showed intermediate susceptibility to streptomycin. On the other hand, among 32 abattoir workers interviewed to assess their knowledge, attitude and practice towards food safety. Almost all abattoir workers were interviewed. In general, PCR and conventional methods used for the detection of the organism highlight the potential threat to public health. The abattoir workers need to be aware about the pathogen and should follow appropriate practices to prevent contamination of meat intended for human consumption.