Objective：To isolate and purify novel Pseudomonas aeruginosa（P. aeruginosa）bacteriophages. Methods： Bacteriophages were isolated from Nanjing Medical University campus Tianyuan lake，followed by genome sequencing and biological characterization of purified phages. In vitro antibacterial assays，including growth inhibition curves and biofilm eradication experiments，were conducted. Results：A novel P. aeruginosa phage named PAPX was successfully isolated. Genomic analysis revealed 98% identity with Pseudomonas phage Epa12，classifying it within the Pbunavirus genus. Transmission electron microscopy demonstrated its myoviral morphology featuring an icosahedral head（∼90 nm diameter）and a contractile tail（∼50 nm length，26 nm diameter）. Biological characterization showed a strong lytic activity against 17 of 24 clinical strains. In vitro experiments confirmed its ability to suppress bacterial growth during early proliferation phase and effectively eradicate P. aeruginosa biofilms. Conclusion：The successful isolation and characterization of phage PAPX，validated through biological profiling and in vitro assays，demonstrates its promising clinical potential for treating P. aeruginosa infections，particularly in combating antibiotic - resistant strains and biofilm -associated infections.