[Background]Escherichia coli and Salmonella are the main pathogens in broiler breeding, which can cause huge economic losses, and their antibiotic resistance shouldn’t be ignored. [Objective] To isolate and identify pathogens from dead chickens, and analyze their antibiotic resistance characteristics. [Methods] The target strains were isolated and purified by conventional method. Biochemical tests, 16S rRNA gene sequence analysis and growth curve verification were used for identification. The drug susceptibility test was carried out by disk diffusion method, and the drug resistance rate and multiple drug resistance were analyzed. [Results] A total of 10 strains of E. coli and 10 strains of Salmonella were isolated, including 4 enterica subspecies, 1 arizonae subspecies and 5 Salmonella bongori. E. coli isolates were only sensitive to carbapenems and resistant to other antibiotics. While Salmonella isolates were sensitive to carbapenems and showed different degree of resistance to other antibiotics. All the isolates were multi-drug resistant. [Conclusion] The pathogens were isolated and their antibiotic resistance were determined, which provided reference for the prevention and cure of E. coli and Salmonella in white feather broiler breeding.