Objective To understand the clinical distribution characteristics and drug-resistance genotypes of carbapenem-resistant Pseudomonas aeruginosa (CRPA) in a tertiary hospital in Shanghai, so as to guide the rational use of antibiotics, reduce bacterial resistance and control nosocomial infections. Methods A total of 94 consecutive and non-redundant CRPA strains isolated from clinical specimens were collected in The First Affiliated Hospital of Naval Medical University between Jan. 1, and Dec. 31, 2019. The distribution of CRPA across departments and sample sources were analyzed. Antimicrobial susceptibility testing against 13 antibiotics was determined using the VITEK 2 Compact automated microbiology analyzer and the Kirby-Bauer disk diffusion method. Polymerase chain reaction (PCR) was employed to detect genes encoding extended-spectrum β-lactamase resistance gene, carbapenemase resistance gene, and porin resistance gene. Homology analysis of selected isolates was performed using kSNP3, a k-mer-based software, for single nucleotide polymorphism (SNP) analysis. An evolutionary tree was constructed to assess genetic relationships. Additionally, multilocus sequence typing (MLST) was performed using the Pseudomonas aeruginosa typing scheme from the PubMLST database. Results Among the 94 CRPA isolates, high resistance rates were observed for carbapenems, including imipenem (92.6%, 87 strains) and meropenem (87.2%, 82 strains). In contrast, low resistance rates were detected for aminoglycosides, such as amikacin (10.6%, 10 strains), gentamicin (20.2%, 19 strains), and tobramycin (20.2%, 19 strains). The top 3 departments in terms of isolate distribution were the Emergency Intensive Care Unit (9.6%, 9 strains), Department of Cerebrovascular Surgery (8.5%, 8 strains), and Department of Respiratory Medicine (8.5%, 8 strains). PCR analysis of 94 CRPA strains detected outer membrane protein D2 (OprD2) gene deletion in 47 (50.0%) strains, 13 (13.8%) strains were positive for bla_KPC, 4 (4.3%) strains for bla_VIM, 2 (2.1%) strains for bla_IMP, 1 (1.1%) strain for bla_NDM, 12 (12.8%) strains for bla_TEM, 4 (4.3%) strains for bla_PER, and 2 (2.1%) strains for bla_GES, while bla_OXA-48, bla_BIC, bla_SIM, bla_VEB, and bla_SHV were not detected. MLST identified 36 different sequence types (STs), with ST463 and ST274 being the most common, and 2 new ST (ST4023 and ST4024) were identified for the first time. Conclusion CRPA strains carry multiple resistance genes and exhibit concurrent resistance to several commonly used clinical antibiotics. The resistance is primarily associated with the presence of bla_KPC, bla_VIM and bla_TEM genes and the deletion of OprD2 gene. Clinical monitoring of CRPA should be strengthened, and rational use of antimicrobial agents is essential to control its spread within the hospital.