OBJECTIVE: To provide reference for rational selection of antibiotics against non-fermentative Gram-negative bacilli in clinic. METHODS: Etiological data of clinical isolated Pseudomonas aeruginosa (PA), Acinetobacter baumanii (AB) and Stenotrophomonas maltophilia (SM) were collected from the Affiliated Hospital of Qingdao University (called “our hospital” for short) during Jan. 2004-Dec. 2016. Drug resistance of them to commonly used antibiotics was analyzed retrospectively. RESULTS: Totally 15 587 strains of PA, 7 446 strains of AB and 2 950 strains of SM were detected. Resistance rates of PA to commonly used antibiotics fluctuated but were in a decreasing tendency. Except for imipenem, resistance rates of PA to commonly used antibiotics decreased significantly, and resistance rates of PA to amikacin and gentamicin decreased to 4.60% and 7.48%, respectively. Resistance rates of AB to most commonly used antibiotics were more than 40%, but it was sensitive to tigecycline (drug resistance of 0-4.03%). Resistance rates of SM to cefoperazone sodium and sulbactam sodium increased from 3.03% in 2004 to 39.01% in 2016, but it was sensitive to sulfamethoxazole, minocycline and levofloxacin. CONCLUSIONS: Non-fermentative Gram- negative bacilli detected in our hospital are mainly PA. Resistance rate of PA to most of the antibiotics is declining; drug resistance of AB is severe; resistance rates of SM to cefoperazone sodium and sulbactam sodium show increasing tendency. Above 3 non-fermentative Gram-negative bacilli are sensitive to amikacin, tegocycline and minocycline. Clinical selection should be based on the results of drug sensitivity test.