Abstract: OBJECTIVE To explore the pathogen distribution and drug resistance of the main pathogens in cholesteatoma and non-cholesteatoma chronic suppurative otitis media before and after surgery. METHODS A total of 170 cases of patients with chronic suppurative otitis media in otolaryngology of Wenzhou Integrative Medicine Hospital of Zhejiang Traditional Chinese Medicine University from Jan. 2008 to Dec. 2015 were selected as research subjects, and were divided into group A (cholesteatoma) with 90 cases and group B (non-cholesteatoma) with 80 cases. The preoperative, intraoperative and postoperative tissue or secretions of the two groups of patients were given pathogen culture and sensitivity test. RESULTS There were 10 strains of fungi, 60 strains of gram-positive bacteria and 74 strains of gram-negative bacteria in group A, and 6 strains of fungi, 59 strains of gram- positive bacteria and 29 strains of gram-negative bacteria in group A before, during and after surgery. The difference of pathogen culture positive rate before and during surgery in group A was not significant. The pathogen culture positive rate before surgery was higher than that of during surgery in group B (P<0.05). The difference of pathogen culture positive rate before surgery between group A and group B was not significant. The pathogen culture positive rate of group A was higher than that of group B after surgery (P<0.05). The 84 strains of Staphylococcus aureus were sensitive to vancomycin, nitrofurantoin, linezolid, and tigecycline, and had high resistant rates to erythromycin and penicillin G, accounting for 75.0% and 86.9%. The 55 strains of Pseudomonas aeromonas were sensitive to cefepime and amikacin, and had the resistant rates of 100% to ceftriaxone, cefotetan, ampicillin, cefuroxime, cefazolin, cotrimoxazole, and nitrofurantoin. CONCLUSION The pathogens of chronic suppurative otitis media before and during surgery are not exactly the same. The pathogens in patients with cholesteatoma has higher detection rate. Different pathogens have different susceptibility test results.