Introduction: Umbilical cord is the first source of blood from the neonate. The clinical signs associated with sepsis are frequently non specific and subtle in the neonates making the diagnosis of infection difficult. Umbilical cord blood does not involve pain infliction, avoids iatrogenic blood loss and procedural complications. Aim: To evaluate the utility of Umbilical Cord Blood Culture (UCBC) in the diagnosis of Early Onset Sepsis (EOS). Materials and Methods: The present study was a crosssectional study carried out in neonatology unit of Department of Paediatrics, Rural Medical College, Loni, Maharashtra, India, for the period of mid-March 2021 to mid-September 2021. Neonates delivered in Rural Medical College to the mothers having the risk factors for EOS were included in this study. Informed consent was taken prior to start of study. Thus, 68 samples were collected and studied. UCBC was collected with all aseptic precautions immediately after the umbilical cord was cut after the birth of the baby, venous blood sample was collected within one hour of birth. The data was presented as count and percentages. Results: Majority were female neonates 40 (58.8%). On analysing maternal risk factors it was seen 8.8% had previous low birth weight and 13.2% had Rh negative status. Analysis for presence of risk factors for sepsis majority 79.4% were multiple times examined per vaginally, followed by 36.8% had foul smelling liquor, 11.8% had febrile illness and 5.8% had birth asphyxia. On umbilical cord culture and sensitivity, most common microorganism identified was Staphylococcus aureus 8 (11.8%), followed by 8.8% Pseudomonas species. So, present study shows that 35.3% (24 cases) had positive culture reports using utility of UCBC in 68 patients. Conclusion: Study concludes that umbilical cord blood sampling and culture can be used as a tool for diagnosing bacterial sepsis in neonates especially the high-risk neonates.