In recent years, multidrug resistant virulent strains of E. coli are implicated in broiler cases of high mortalities causing significant economic losses in poultry industry. The current study aimed to determine the prevalence of E. coli isolates and to discuss the distribution of some virulence genes among multidrug resistance E. coli isolates. Bacteriological examination of 242 samples collected from different sources in Sharkia province, Egypt revealed a low recovery rate of E. coli isolates (20.2%). Serotyping of the representative E. coli isolates revealed 8 different serotypes with a dominance of O26:K60 and O114:K90 serotypes (20.4% for each one). Antimicrobial susceptiblility testing of E. coli isolates demonstrated that all the isolates were multidrug resistant and all E. coli isolates were resistant to more than 11 antimicrobial agents. Moreover, the highest resistance rates were recorded against trimethoprim sulfamethoxazole, amoxicillin clavulanic acid and erythromycin (100%), while the least resistance rate was detected against doxycycline (51.02%). With regard to the molecular detection of some virulence (iss and pap < /em>C) and resistance genes (blaTEM and tetB) on the extracted plasmid of 11 E. coli isolates were tested. Overall, these data indicated that the dissemination of resistance is associated with genetic mobile element such as plasmids that may also carry virulence determinants.