To study the effect of different antibiotic use strategies on infection in neonates with premature rupture of membranes and the high-risk factors for neonatal infection. A retrospective analysis was performed for the clinical data of 223 neonates with premature rupture of membranes. According to the antibiotic use strategies, they were classified into two groups: study group (n=95) and control group (n=128). The neonates in the study group were given antibiotics based on risk assessment and infection screening, and those in the control group were given antibiotics based on risk assessment alone after admission. General status and prognosis were compared between the two groups. According to the presence or absence of infection, the neonates were classified into two groups: infection group (n=32) and non-infection group (n=191). The clinical data were compared between the two groups. A logistic regression model was used to investigate the high-risk factors for infection in neonates with premature rupture of membranes. Compared with the control group, the study group had a significantly longer time of premature rupture of membranes, a significantly higher rate of cesarean section, and a significantly lower antibiotic use rate at admission and a significantly lower total antibiotic use rate (P<0.05). The study group also had a significantly higher incidence rate of infection and a significantly lower incidence rate of sepsis (P<0.05). There were no significant differences in the incidence rates of other complications between the two groups (P>0.05). Compared with the non-infection group, the infection group had a significantly lower gestational age, a significantly longer time of premature rupture of membranes, a significantly higher rate of cesarean section, and significantly higher levels of C-reactive protein (CRP) and procalcitonin on admission and during reexamination (P<0.05), with fever as the most common symptom. The logistic regression analysis showed that preterm birth and cesarean section were high-risk factors for infection in neonates with premature rupture of membranes (P<0.05). Strict adherence to the indications for antibiotic use in neonates with premature rupture of membranes does not increase the incidence rate of complications. Neonates with premature rupture of membranes, especially preterm infants and infants delivered by cesarean section, should be closely observed for the change in disease conditions, and infection indices including CRP should be reexamined in case of fever and antibiotics should be used to prevent serious infection.