Objective To analyze the perinatal risk factors and pathogen distribution of neonatal early-onset septicemia(EOS). Methods 168 neonates with EOS in the neonatal intensive care unit (NICU) of a hospital from 2016 to 2019 were selected as infection group, and 168 neonates without septicemia in NICU were randomly selected as control group, data about general condition and perinatal infection of neonates' mothers, as well as general condition, invasive procedure and complications of neonates were collected and compared. Results Neonates with EOS were mainly extremely premature neonates (n=65, 38.69%), neonates in control group were mainly full-term (n=75, 44.65%). In EOS group, most were extremely low birth weight neonates (n=67, 39.88%) and low birth weight neonates (n=54, 32.14%), while in control group, majority were normal weight neonates (n=98, 58.33%). Univariate analysis showed that mothers' gestational age, the number of induced abortion, placental abnormalities, amniotic fluid abnormalities, premature rupture of membrane, prenatal fever, as well as neonatal gender, gestational age, birth weight, and moist lung were influencing factors for neonatal EOS (all P < 0.05). Multivariate logistic regression analysis showed that mothers' gestational age>35 years, induced abortion, premature rupture of membrane, prenatal fever, neonatal birth weight lower than normal, and moist lung were independent risk factors for EOS. The main pathogens of neonatal EOS were Escherichia coli, Staphylococcus epidermidis and Klebsiella pneumoniae. Conclusion Early intervention should be carried out according to the risk factors for neonatal EOS, the appropriate treatment scheme should be selected according to the main pathogens.