Objective To explore the application effect of information-based transparent supervision intervention model in the prevention and control of multidrug-resistant organism (MDRO) healthcare-associated infection(HAI). Methods 46 873 inpatients in a hospital From January to December 2019 were as pre-intervention group and 45 217 inpatients from January to December 2020 were as post-intervention group. Transparent supervision was implemented by intervention group for patients who were isolated MDRO and early warned through daily HAI real-time monitoring system. With the help of information, infection rate and isolation rate of MDRO, qualified rate of ATP detection on hand hygiene and environmental object surface cleaning and disinfection, as well as implementation rate of MDRO core prevention and control measures were transparently opened, effect before and after transparent supervision intervention was compared. Results After information-based transparent supervision intervention, infection rate of MDRO decreased from 0.96% to 0.51% (P < 0.001), isolation rate of MDRO decreased from 56.04% to 40.79% (P < 0.001), hand hygiene compliance rate increased from 53.31% to 70.25% (P < 0.001); qualified rate of environmental object surface cleaning and disinfection increased from 51.89% to 91.76% (P < 0.001); after intervention, detection rate of pathogenic organism before the therapeutic use of antimicrobial agents increased gra-dually (P < 0.001), utilization rate of special grade antimicrobial agents generally decreased (P < 0.05). Conclusion With the help of information-based real-time monitoring system, timely early warning MDRO infection cases, strengthening transparent supervision on the implementation of MDRO core prevention and control measures as well as applying information platform to promote the process controlling of rational use of special-grade antimicrobial agents can effectively reduce MDRO cross infection in hospital and reduce the outbreak of MDRO infection.