ObjectiveTo understand the current situation of multidrugresistant organism (MDRO) infection in hospitalized patients in a tertiary firstclass hospital, as well as efficacy of bundle intervention measures on prevention and control of infection. MethodsHospitalized patients who were infected with MDROs in this hospital during 2012-2014 were monitored and conducted bundle intervention. ResultsIn 2012-2014, a total of 1 909 MDRO strains were isolated. The isolation rates of MDROs in 2012-2014 were 30.69%, 31.04%, and 29.40% respectively，bedside intervention rates were 81.02%, 92.05%, and 94.23% respectively，implementation rates of clinical isolation were 69.97%, 82.98%, and 93.04% respectively; MDRO isolation rates during 2012-2014 were not significantly different（P＞0.05）；bedside intervention rates and implementation rates of clinical isolation were both significantly different（both P＜0.05）. The top 3 isolated MDROs in 2012-2014 were extendedspectrum βlactamases(ESBLs)producing Escherichia coli, multidrugresistant Acinetobacter baumannii, and ESBLsproducing Klebsiella pneumoniae. ConclusionBundle intervention measures, such as multidepartment collaboration and strengthened onsite supervision, can promote more standardized management of healthcareassociated infection, enhance the implementation rate of clinical isolation, and effectively prevent and control the spread of MDRO in hospital.