METHODS: Isolated phage vB_Kpn_HF0522 and phage morphology were observed using transmission electron microscopy. Analysis of vB_Kpn_HF0522 characteristics, including optimal multiplicity of infection (MOI), one-step growth curve, host range, stability in different environments, and adsorption capacity. The phage genomic sequence was analyzed to explore evolutionary relationships. The effect of phage vB_Kpn_HF0522 on biofilms was assessed using crystal violet staining assay. The RESULTS: Phage was isolated from hospital sewage for experimental studies. Genome analysis revealed that vB_Kpn_HF0522 is a double-stranded linear DNA virus. Biological characterization demonstrated that this phage specifically targets serotype K1 CONCLUSION: These findings indicate that vB_Kpn_HF0522 has significant potential for treating specific bacterial infections, and may serve as an antimicrobial agent for research and clinical anti-infective therapy.