Neonatal sepsis, a major newborn killer worldwide exhibits wide spectrum of clinical manifestations. Epidemiology and aetiological organisms vary with geographical area and time. Objective of the study was to study clinical characteristics, epidemiology, bacterial aetiology and drug sensitivity pattern of isolated organisms in Neonatal Intensive Care Unit (NICU), Dhaka Medical College Hospital (DMCH), Dhaka, Bangladesh. This observational cross sectional study was carried out in NICU, DMCH from January 2014 to June 2015. The inclusion criteria were newborns having features of sepsis at admission or developing such features afterwards when admitted for other indications. A blood sample was collected aseptically immediately after clinical diagnosis and was sent for relevant laboratory investigations. Sample for culture sensitivity was inoculated bedside to culture bottle and sent to department of microbiology. The newborns were followed up till hospital discharge or death. All information regarding history, laboratory findings and follow up were recorded in a structured questionnaire. Of the 200 neonates, 59% were diagnosed as having late onset sepsis (LONS). Premature and low birth weight (LBW) babies mostly suffered from LONS. Respiratory distress, tachycardia, lethargy were the predominant symptoms in both early and late sepsis. Blood culture yielded growth in 55% of the septic newborns. Klebsiella pneumoniae was the predominant organism in both early and late sepsis. Most of the Gram negative bacteria were sensitive to colistin, meropenem and imipenem. Case fatality was 24.39% and 34.74% in early and late sepsis respectively.