Aim To develop a novel in vitro urethra model and use it to determine if insertion of an intermittent urinary catheter (IC) displaces pathogenic bacteria from the urethral meatus along the urethra. Methods Displacement of microbial growth after catheter insertion was assessed using a novel in vitro urethra model. The in vitro urethra model utilized chromogenic agar and was inoculated with bacteria at one side of the artificial urethra channel, to act as a contaminated urethral meatus, before an IC was inserted into the channel. Three ICs types were used to validate the in vitro urethra model and methodology. Results When compared to the bacterial growth control, a significant difference in bacterial growth was found after insertion of the uncoated (P <= 0 center dot 001) and hydrophilic coated (P <= 0 center dot 009) catheters; no significant difference when a prototype catheter was inserted into the in vitro urethra model with either bacterial species tested (P >= 0 center dot 423). Conclusion The results presented support the hypothesis that a single catheter insertion can initiate a catheter-associated urinary tract infection. Significance and Impact of the Study The in vitro urethra model and associated methodology were found to be reliable and reproducible (P >= 0 center dot 265) providing new research tool for the development and validation of emerging technologies in urological healthcare.