In present study, aim was to determine the bacteriological profile in Neonatal Sepsis and antimicrobial sensitivity of the organisms in the Neonatal Intensive Care Unit (NICU) of Tertiary care Hospital and to identify risk factors for neonatal sepsis. This study included records of all cases of clinically suspected septicemia in neonates admitted to the NICU from September 2015 to August 2016 (n=101). Result: The risk of sepsis in birth asphyxia and gestational age was about 2 times higher as compared to the control group and p- value was significant at 0.05 levels. Out of the total 95 cases having perinatal fever (n = 95, 94%), 34 (34%) were culture positive and 61 (60%) were culture negative, which was statistically significant. Conclusion: Klebseilla (Gm – ve) (23%) and Staphylococcus aureus (Gm + ve) (15.3%) were the most common organisms causing neonatal sepsis in our settings. Ampicillin and Gentamicin for 1st line, Ciprofloxacin and Piperacillin and Tazobactam for 2nd line, Meropenum and Vancomycin for 3rd line drugs for empirical antibiotic therapy.