Abstract: Objective To analyze risk factors of multiple drug-resistant infections in neonatal intensive care unit (NICU). Methods The clinical data from 284 hospitalized pediatric patients were retrospectively analyzed from June 2011 to July 2015. The differences between 59 cases with multiple drug-resistant infections and 225 cases with non-multiple drug-resistant infections were compared and analyzed by logistic regression. Results All of 284 cases were single birth. Fifty-nine cases (13.13±9.03 days old) had multiple drug-resistant infections, in which 42 were males and 17 were females. Two hundred and twenty-five cases (14.21±8.34 days old) had non-multiple drug-resistant infections, in which 175 cases of males and 50 cases of females.. Single factor analysis showed that 8 factors, including gestational age, birth weight, days in hospital, Apgar score at birth, mechanical ventilation, parenteral nutrition, and the categories and duration of use of antimicrobial agents, were the risk factors of multiple drug-resistant infections (P < 0.05). Logistic regression analysis showed that the birth weight and the categories and duration of use of antimicrobial agents were the dominant factors that caused multiple drug-resistant infections in NICU (P < 0.05). Conclusions Effective prevention and control measures should be taken to reduce the multiple drug-resistant infection in NICU.