Abstract: OBJECTIVE To study the effect of comprehensive interventions such as active monitoring on the prevention and controlling of nosocomial infections in neonatal ward. METHODS A retrospective survey collected data on 5 305 cases of infection patientsin neonatal wards of the hospital before the implementation of comprehensive intervention from Jan. 1, 2014 to Dec. 31, 2015, and 8 456 cases of patients underwent active monitoring and comprehensive intervention measures including standard prevention for neonates with high risk of infection, contact and isolation prescription for those diagnosed multidrug-resistant bacteria, supervision, supervision of the implementation of isolation measures and hand hygiene compliance from Jan. 1,2016 to Dec. 31, 2018. Data were collected prospectively after the intervention, and the changes in the incidence of nosocomial infection, detection of drug-resistant bacteria, and compliance with hand hygiene before and after the intervention were analyzed and compared. RESULTS The incidence of nosocomial infection rate decreased from 1.96%（104/5 305） to 1.5%（127/8 456）（P=0.042）, hospital infection rate of MRSA and CRE decreased from 0.17%（9/5 305） and 0.21%（11/5 305）, respectively to 0.05%（4/8 456） and 0.08%（7/8 456）, respectively（P＜0.05）.Isolation rate increased from 52.38%（22/42） to 90.00%（27/30）（P=0.001）, and hand hygiene compliance rate increased from 90.07%（363/403） to 94.05%（1 471/1 564）（P=0.005）. CONCLUSIONS Comprehensive intervention measures such as active monitoring, standard prevention, contact and isolation prescription, for patients diagnosed with multidrug-resistant bacteria, supervison of the implementation of quarantine measures and compliance with hand hygiene can effectively reduce the incidence of nosocomial infection in neonatal patients and spread of multidrug resistant nosocomail infection.