Neonatal sepsis is the leading cause of infant mortality and morbidity on a global scale. This was a retrospective study that aimed to assess and to evaluate the bacteriological profile, antibiotic resistance and sensitivity pattern in neonates with sepsis in a tertiary care hospital. The neonatal cases with sepsis were selected by inclusion and exclusion criteria. All neonates with a clinical suspicion of sepsis with a positive blood culture were identified. Patient demographics, clinical details, maternal risk factors and laboratory data including bacteriological profiles. Antimicrobial susceptibilities were analyzed and recorded. Out of 110 neonatal cases, the most predominant bacteria isolated were Gram-negative organisms, they were Klebsiella pneumonia (n=27, 41.5%), Acinetobacter sp (n=14, 21.5%) and the majority of Gram-positive bacterial isolate was Staphylococcus aureus (n=9, 20%). The antibiotics Amikacin (79.3%), Gentamycin (76.1%), Imipenem (74.6%), Meropenem (71.4%) and Ciprofloxacin (69.8%) were sensitive and Ampicillin (87.3%), Amoxiclav. (71.4%), Cefopodoxime (66.6%) and Cefuroxime (61.9%) were resistant to Gramnegative bacteria. The antibiotics Amikacin (88.8%), Gentamycin (82.2%) and Chloramphenicol (86.6%) were sensitive and the antibiotics Ampicillin (77.7%), Cefixime (73.3%) were resistant to Gram-positive bacteria. The research aids us in concluding that regular antibiotic surveillance is essential at the study site to reduce antibiotic resistance.