Abstract: Objective To explore the pathogenic bacteria distribution and drug resistance of otitis media in preschool children. Methods Children with otitis media during January 2012 to December 2015 were selected. Ear discharge was taken for common pathogenic bacteria isolation and culture. The drug sensitivity was tested. Results In 200 strains of pathogenic bacteria isolated, 156 (78.0%) strains were Gram-positive cocci, mainly being Streptococcus pneumoniae, Staphylococcus aureus; 38 (19.0%) strains were Gram-negative bacilli, mainly being Pseudomonas aeruginosa; 6 (3.0%) strains were monilia. Streptococcus pneumoniae had high rate of resistance to erythromycin, tetracycline, clindamycin and sulfamethoxazole, and low rate of resistance to penicillin. Staphylococcus aureus had high rate of resistance to penicillin, and had not found resistance to vancomycin, linezolid and daptomycin. In routine monitoring of 12 kinds of antimicrobial agents, the overall drug resistance of Pseudomonas aeruginosa was low, and the resistance rates of each antimicrobial agent was less than 32%. Conclusion The common pathogens of otitis media in preschool children were Streptococcus pneumoniae, Staphylococcus aureus, and Pseudomonas aeruginosa. However, different bacteria had different drug resistance. Thus, rational use of antibacterial drugs is the key to control infections.