To understand the distribution of Escherichia coli in poultry and to reveal the drug resistance and molecular epidemic and characteristics of isolates from Shandong Province, and to provide science basis for the risk assessment.Methods A total of 42 E.coli isolates were analyzed with drug sensitivity test for 13 antimicrobial agents, and analyzed with PFGE and MLST genotyping. Results The PFGE genotyping of 42 isolates yielded 34 PFGE types. MLST analysis of isolates identified 18 STs, and the 7 housekeeping genes had different variation. All isolates were resistant to two or three antimicrobial agents, and the highest resistance was to AM, SXT and NOR, which was 97.62%、92.86% and 90.48%, respectively.Conclusion The resistant spectrum of E.coli was wide and serious, gene was polymorphism, and the distribution had a certain timeliness and regional in parts of Shandong Province.