Background To ensure that new acquired clinics perform sterilization and high-level disinfection (HLD) according to best practice guidelines, a comprehensive competency program was begun by our Infection Prevention (IP) Program. The competency program included a classroom session, reprocessing demonstration, and a subsequent competency checkoff. The program reviewed topics pertaining to point of care cleaning, cleaning, instrument inspection, preparation and packaging, autoclaving, logging, and biological indicators (for sterilization) and other quality control items. Methods IP specialists conducted a detailed inspection and comprehension assessment of the clinics’ sterilization and HLD practices that mirrored the competency program. Baseline IP surveys were completed between 2014-2016 and were compared to phase-one (2016-2017), phase-two (2017-2018) and phase-three surveys (2019-2020) to measure the competency programs impact on reprocessing practices. Only clinics that performed sterilization or HLD during all three surveys were included. For analysis, the sterilization scores were further divided into reprocessing area, point of care/cleaning, preparation/quality, and logging/storage subcategories. HLD scores were separated into reprocessing area, point of care/cleaning, preparation/quality, and after HLD/logging/storage subcategories. Kruskal-Wallis test was used to determine significance for the overall clinic average. Results A total of 38 clinics were included in the sterilization analysis, and 8 clinics in the HLD. The overall clinic scores for sterilization were baseline (65%), phase-one (93%), phase-two (94%) and phase-three (94%) (p-value: Conclusions The competency program improved sterilization and HLD reprocessing in every category assessed and it has sustained for the next five years validating the value of a comprehensive competency program with an education, demonstration, and evaluation component. To ensure that new acquired clinics perform sterilization and high-level disinfection (HLD) according to best practice guidelines, a comprehensive competency program was begun by our Infection Prevention (IP) Program. The competency program included a classroom session, reprocessing demonstration, and a subsequent competency checkoff. The program reviewed topics pertaining to point of care cleaning, cleaning, instrument inspection, preparation and packaging, autoclaving, logging, and biological indicators (for sterilization) and other quality control items. IP specialists conducted a detailed inspection and comprehension assessment of the clinics’ sterilization and HLD practices that mirrored the competency program. Baseline IP surveys were completed between 2014-2016 and were compared to phase-one (2016-2017), phase-two (2017-2018) and phase-three surveys (2019-2020) to measure the competency programs impact on reprocessing practices. Only clinics that performed sterilization or HLD during all three surveys were included. For analysis, the sterilization scores were further divided into reprocessing area, point of care/cleaning, preparation/quality, and logging/storage subcategories. HLD scores were separated into reprocessing area, point of care/cleaning, preparation/quality, and after HLD/logging/storage subcategories. Kruskal-Wallis test was used to determine significance for the overall clinic average. A total of 38 clinics were included in the sterilization analysis, and 8 clinics in the HLD. The overall clinic scores for sterilization were baseline (65%), phase-one (93%), phase-two (94%) and phase-three (94%) (p-value: The competency program improved sterilization and HLD reprocessing in every category assessed and it has sustained for the next five years validating the value of a comprehensive competency program with an education, demonstration, and evaluation component.