Objective To investigate the distribution of multidrug-resistant organisms (MDRO) and other pathogens in the patients with chronic suppurative otitis media(CSOM) to provide evidence for prevention and treatment of CSOM. Methods: A total of 224 patients with CSOM were enrolled from January 2016 to March 2020. The samples of ear secretion were obtained for bacterial culture and isolating pathogen, and the test of drug sensitivity was conducted. Results: The samples from 177 patients (78.57%) were positive in bacterial culture and a total of 215 strains were detected, inclu-ding 72 strains of gram-positive bacteria(72/215, 33.49%), 19 strains of gram-negative bacteria (19/215, 8.84%), 124 strains of fungi(124/215, 57.67%). Among 215 strains, 36 strains of MDRO(36/215, 16.74%) were isolated, including 20 strains of Staphylococcus aureus(20/36, 55.56%), 12 strains of coagulase negative Staphylococcus(CoNS)(12/36, 33.33%) and 2 strains of Pseudomonasaeruginosa (2/36, 5.56%). Staphylococcus aureus and CoNS had highest resistant rate to penicillin (95.10% and 64.29%), while Pseudomonas aeruginosa had highest resistant rate to levofloxacin(62.50%). The statistical analysis revealed that empirical antibiotic treatment was an independent risk factor of MDRO infection (P<0.05), and the fungal infectionin patients had no relationship with empirical antibiotic treatment, gender or age. Conclusions: Fungal infection is the main cause for local CSOM. The bacterial culture combined with the test of drug sensitivity are essential for treatment of CSOM, and empirical antibiotic treatment should be avoided to use to reduce MDRO infection.