Clinically suspected 77 cases of neonatal septicemia admitted to the pediatric ward of BP Koirala Institute of Health Sciences, Dharan, Nepal, were evaluated by blood culture. The blood culture was positive for bacterial growth in 46 (59.7%) cases. Gram-negative bacteria were isolated in 33 (71.7%) cases, the most common being Escherichia coli, followed by Klebsiella spp, Enterobacter spp. Staphylococcus aureus was the most common gram-positive bacteria. The bacterial isolates were most sensitive to gentamicin (74.5%) and ciprofloxacin (74.5%) followed by chloramphenicol (59.9%). They were most resistant (78.8%) to ampicillin. This study for the first time underlines the pattern of bacterial isolates and their sensitivity pattern to antibiotics in this part of Nepal.