Objective To understand the distribution of clinical isolates of Pseudomonas aeruginosa (PA), prevalence of multidrug-resistant PA (MDR-PA), extensively drug resistant-PA (XDR-PA), and carbapenem-resistant PA (CRPA) in a tertiary first-class hospital in Shanghai, so as to provide evidence for rational use of antimicrobial agents. Methods Specimen information and antimicrobial susceptibility data of PA isolated from cultured specimens of clinical departments from 2014 to 2017 were collected, sources of specimens and department of PA, relationship between PA and age as well as sex of patients were analyzed, isolation rates of MDR-PA, XDR-PA and CRPA were calculated. Results A total of 1 218 PA strains were isolated in the past four years, number of isolated PA increased year by year; strains were mainly isolated from sputum (75.0%), followed by urine (10.6%). PA was most common in department of geriatric medicine (44.9%), followed by intensive care unit (8.9%) and respiratory department (8.9%), PA was mostly isolated from people over 60 years old and mainly male. Among PA strains, percen-tage of MDR-PA was 30.5%, but XDR-PA was only 2.5%, while isolation rate of CRPA was up to 38.8%, which showed an increasing trend year by year. Specimen sources and department distribution of MDR-PR, XDR-PA and CRPA were similar to those of PA, that is, sputum and department of geriatrics medicine. Conclusion Isolation rate of MDR-PA and CRPA in this hospital is high, strains are mainly from sputum and department of geriatric medicine, prevention and control measures of healthcare-associated infection(HAI) should be strengthened accor-ding to different departments and populations, management of clinical use of antimicrobial agents should be strengthened to prevent and control the occurrence and prevalence of MDR-PA and CRPA.