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Objective: This study was to establish a mice model for catheter-related biofilm infection suitable to bioluminescence imaging (BLI).Methods: Biofilms of Pseudomonas aeruginosa (P.aeruginosa) Xen5 to be grown on catheter disks in vitro and implanted into mice to induce catheter-related biofilm infections in vivo.Biofilm formation in vitro and in vivo was real-time monitored during the study period using BLI.The images were analysed, and the relationship between integrated brightness (IB) and viable bacterial count (VBC) was compared.In addition, we monitored the change in white blood cells (WBCs) during the biofilm infection.Results: The levels of IB and VBC in biofilm disks in vitro were highest at 24 h after P.aeruginosa Xen5 inoculation, IB of biofilm in implanted mice model was increased until 24 h.A good relationship was observed between IB and VBC in vitro by linear regression analysis.The actual number of VBC in the infected catheter disks at day 7 of implantation was very close to the estimated number according to the formula of linear regression from in vitro study.The number of WBCs in the control group was gradually increased from day 1 to day 5 but restored to normal level after that, while their number in the infection group was gradually increased during the study period.The number of WBCs on day 7 was significantly higher in the infection group than control group.Conclusion: Bioluminescence imaging is a simple, fast and sensitive method to measure the catheter biofilm infection in mice.