Objective　To explore the clinical characteristics, diagnosis and treatment of early onset sepsis caused by Listeria monocytogenes in neonates. Methods　The clinical data of early onset sepsis with positive blood culture of Listeria monocytogenes in 12 neonates admitted to NICU from January 2011 to June 2018 were retrospectively analyzed. Results　Among the 12 cases, 8 were males, 4 were females, and 10 were premature. The main clinical manifestations were abnormal body temperature, shortness of breath, distress or apnea, convulsions, rashes and pulmonary hemorrhage. Blood culture results showed Listeria, and Listeria was also found in cerebrospinal fluid culture in one case. The antibiotics, including meropenem, penicillins and vancomycin, were used alone or in combination, and 4 cases were improved and 8 cases died. Conclusion　Neonatal sepsis caused Listeria monocytogenes is a very serious infectious disease, which is characterized by acute onset, severe infection and high mortality.