The research work was undertaken for detection and determination of antibiogram profile of Escherichia coli (E. coli) that produce Shiga toxin from apparently healthy broilers (n=8) from different commercial farms of Jessore, Bangladesh. Broiler cloacal swabs (n=8) were collected by inserting the sterile swab stick in the cloacae of broiler and inoculated into peptone water for enrichment for 24 hours at 37°C. Enriched culture was streaked onto Eosin Methylene Blue (EMB) agar for isolation of pure bacterial culture. Then pure bacterial culture was streaked onto Mac-Conkey (MC) agar to identify Gram negative bacteria. Cultural method, Gram staining, biochemical reaction and polymerase chain reaction technique were used to identify the bacteria. The antibiogram profiles of bacteria were investigated against 6 commonly used antibiotics (Ciprofloxacin, Ampicillin, Colistinsulphate, Erythromycin, Neomycin and Penicillin) by disc diffusion method. Five E. coli isolates were identified and Shiga toxin producing E. coli (STEC) was detected by amplifying 372-bp fragment of Stx2 gene in polymerase chain reaction (PCR) assay. The prevalence of the Shiga toxin generating E. coli (STEC) in broiler cloacal swab was 62.5%. All isolates (100%) were resistant to Ampicillin, Colistinsulphate, Erythromycin, Neomycin and Penicillin and sensitive to Ciprofloxacin. The findings of this research strongly imply that broiler harbor multidrug resistant and Shiga toxin producing E. coli (STEC) which may cause public health problem if enter into human food chain.