This cross-sectional descriptive study was conducted at the neonatal intensive care unit (NICU) in the department of Neonatology, Mymensingh Medical College Hospital (MMCH), Mymensingh, Bangladesh from July 2017 to December 2017 to determine antimicrobial sensitivity pattern in neonatal sepsis. Ninety four neonates (0-28 days) who were admitted in NICU with suspected sepsis were included in this study by purposive sampling technique. After admission written informed consent from parents or guardians obtained and then septic screening along with blood culture and antimicrobial sensitivity was done. All data were compiled, tabulated and then analyzed by SPSS version 21.0. Among 94 cases, 68(72.3%) were preterm and 26(23.4%) were term. There was male predominance and male female ratio was 1.9:1. Most of the patient admitted within 72 hours of birth. Most (84%) had low birth weight (<2500gm). Pre-mature onset of labour, pre-mature rupture of membrane >18 hours, vaginal route of delivery, instrumental resuscitation, pre-lacteal feeding, bottle feeding were the major perinatal risk factors in this study. Early onset sepsis (76.6%) was most prevalent in this study. Blood culture yielded positive growth in 20(21.3%) cases. Among them, Klebsiella was found in 7(35%). E. coli in 6(30%), Acinetobacter was in 3(15%), Staphylococcus aureus in 2(10%) cases. Pseudomonas and Enterobacter were found in rest 2(10%) of the cases. Gram negative bacteria were found in 18(90%) cases. Klebsiella was sensitive to Imipenem (85.7%), Colistin (85.7%) and Ciprofloxacin (77.5%). Sensitivity of E. coli was Imipenem (100%), Colistin (100%), Amikacin (66.7%), Ciprofloxacin (66.7%), Netilmicin (66.7%) and Gentamicin (50%). Acinatobecter had sensitivity to Netilmicin, Colistin, Imipenem (100%). Staphylococcus was 100% sensitive to Imipenem, Netilmicin and Vancomycin. Pseudomonas was found sensitive to Imipenem (100%), Amikacin (100%), Netilmicin (100%) and Colistin (100%). Enterobacter was found highly sensitive to Ciprofloxacin, Colistin and Imipenem. Almost all organisms were resistant to Ampicillin, Gentamicin, Cefotaxime and Ceftazidime. Based on result it is concluded that Klebsiella pneumoniae and Escherichia coli are the leading cause of neonatal sepsis in this study and most of them resistant to multiple antibiotics. Organisms found more sensitive to Imipenem, Colistin, Ciprofloxacin and Netilmicin.