Context: Neonatal septicaemia is most common cause of morbidity and mortality in NICU in developing countries with emergence of antibiotic resistant organisms. For effective management of neonatal septicaemia cases and to formulate antibiotic policy for NICU, blood culture is most important investigation. Aims: This study was carried out to study clinical, microbial profile and antibiotic sensitivity pattern of neonatal septicaemia at our institute. Settings and Design: Present study was a prospective observational study, carried out in the Department of paediatrics, IGGMC Nagpur. Methods __ampersandsignamp; material: 292 neonates admitted with diagnosis of neonatal septicaemia were included in the study. Diagnostic work up included complete hemogram with peripheral blood smear, CRP, blood culture and sensitivity (C/S) and other relevant investigations according to cases. Statistical analysis: Obtained data was analysed and presented. 95 % confidence interval values were also calculated. Results: Out of 292 cases of neonatal septicaemia blood culture was positive in 107(36.64%). Culture positivity in EOS __ampersandsignamp; LOS were 66(34.74%) __ampersandsignamp; 41(40.20%) respectively. In EOS gram negative organisms (83.33%) were common isolates while in LOS gram positive organisms (46.34%) were predominant. Gram positive were sensitive to vancomycin, linezolid while gram negative were sensitive to imipenam, amikacin, piperacillin + tazobactum. Conclusion: Neonatal septicemia is an important cause of morbidity and mortality. This is due to infection by both Gram positive and gram negative bacteria most of which are multi drug resistant especially in the hospital environment. Acinetobacter spp is one of the emerging causes of neonatal sepsis.