Objective To track and evaluate the improvement efficacy of a 3-year continuous implementation of special action of "Improving the pathogen detection rate before antimicrobial therapy", and provide evidence-based basis for future work. Methods Clinical data of inpatients in a tertiary comprehensive hospital from 2020 to 2023 were collected. The baseline survey result in 2020 was taken as the pre-improvement group, and the continuous implementation of special action improvement goal from 2021 to 2023 was as the post-improvement group. Measures were taken, including improving the information system, establishing a multi-department collaboration mechanism, providing multi-level training and education for all staff, standardizing medical behavior and pathogen detection processes, and strengthening supervision efficiency. Indicators were dynamically tracked and strategies were fo-llowed up promptly. Monitoring and data acquisition were carried out through the hospital infection information system. R 4.1.3 statistical software was adopted to compare the differences between two sets of indicators and the changing trends of data in different years, and the improvement efficacy was evaluated. Results After promoting the improvement goal of 3-year special action, the therapeutic antimicrobials usage rate decreased, presenting a downward trend with years (P<0.001). Pathogen detection rate before antimicrobial therapy increased from 39.38% to 85.40%; blood culture detection rate increased from 14.11% to 49.28%; pathogen detection rates before restricted and special antimicrobial therapy increased from 31.76% and 55.97% to 92.11% and 99.10%, respectively; pathogen detection rate before combined use of key antimicrobial agents increased from 83.09% to 97.74%, all presenting increasing trends year by year (all P<0.001). The detection rate of multidrug-resistant organisms decreased. Detection rates of carbapenem-resistant Enterobacterales (CRE) and methicillin-resistant Staphylococcus aureus (MRSA) presented downward trends (P<0.001). Healthcare-associated infection (HAI) diagnosis-related pathogen detection rate remained above 90%. Consistency rate between specimen collection and infection sites increased from 73.26% to 91.67%, with an increasing trend year by year (P<0.05). The internal medicine department had the lowest consistency rate, while the critical care medicine department had the highest consistency rate. Conclusion Three-year continuous promotion of the special action improvement goal and dynamic evaluation have greatly improved the clinical medical personnel’s capability in judging the indicators and detection timing of pathogen specimens accurately, standardized diagnosis and treatment behavior, and guided the correct and rational use of antimicrobial agents in clinical practice, thus reduced the occurrence of bacterial resistance in hospital.