This study was implemented to isolate, characterize the presence of E coli and study their antibiotic resistance and virulence genes in broiler chickens in Assiut city. A total of 120 samples (liver, heart, yolk sac and lung) were gained from 3 to 35 days old clinically and freshly dead broiler suffering from respiratory manifestation (CRD), omphalitis, septicemia and diarrhea in Assiut Governorates for the detection of pathogenic E coli. Isolation and phenotypic identification of the isolates were performed. Serology and detection of antibiotic sensitivity and resistance were done. Also, detection of genes accountable for virulence (ompA and iroN genes) and antimicrobial resistance were all performed on the samples. Also, resistance genes to antimicrobials (blaTEM, blaVIM and qnrA genes) were detected. E coli was detected and recognized in 31.7 % of the cases. According to the data, 11 of the 38 E coli isolates were identified using serology. The conventional disc diffusion method was used to assess the susceptibility and resistance of the isolated E coli to various antibacterial agents. A total of 81.5 % of isolates have a MAR index exceeding 0.2, whereas 18.5 % have a MAR index not more than 0.2. with an average MDR index of 0.485. Antimicrobial resistance genes were detected in 73.7 % of 19 serologically recognized virulence and antibiotic resistance genes in E. coli isolates such as ompA gene detected in 95%, blaTEM gene detected in 95 %, blaVIM gene detected in 73.7 %, and qnrA gene detected in 31.5 %, but the iroN gene was not detected.