Objective To investigate nosocomial infection rate in Intensive Care Unit (ICU), its risk factors and the pathogenic characteristics of multidrug-resistant bacteria through targeted monitoring, in order to provide scientific references for reducing nosocomial infection. Methods Targeted monitoring was performed on the patients who were admitted to the comprehensive ICU between July 2014 and June 2016. Results Nosocomial infection occurred in 312 of the 4 991 patients. The case infection rate was 6.25%, and case infection rate per day was 19.03‰. After the adjustment, the case infection rate per day was 6.77‰. The ventilator-associated pneumonia infection accounted for 30.78‰; catheter-related bloodstream infection occupied 0.30‰; and catheter-associated urinary tract infection accounted for 0.27‰. The respiratory tract was the major part of nosocomial infection, accounting for 90.38%. Gram-negative bacilli were the major bacteria accounting for 92.74%, in whichAcinetobacter baumannii accounted for 36.29%. Conclusions Through targeted monitoring to keep abreast of the current situation of nosocomial infection in ICU, management and interventions can be targeted. It is an important way to reduce nosocomial infection in ICU.