Background: Neonatal sepsis is defined as a clinical syndrome in an infant 28 days of life or younger, manifested by systemic signs of infection and isolation of a bacterial pathogen from the bloodstream. Neonatal sepsis is caused by Gram-positive and Gram-negative bacteria and candida. Microbial invasion of the bloodstream can have serious consequences such as shock, multi-organ failure, disseminated intravascular coagulation (DIC) and death. This study was conducted to know the prevalence of neonatal septicaemia. Methods: This retrospective observational study was conducted in the Department of Microbiology. The data was collected from the records of the Department for a period of one year (January 2014-December 2014). Results: Among 128 blood samples processed from clinically suspected neonatal septicaemic cases, 45 (35.1%) samples showed growth. Among 45 isolates, 22 were from early onset septicaemia (EOS) and 23 were from Late Onset Septicaemia (LOS). Among 45 culture positives, 32 (71.1%) were from males and 13(28.9%) were from females, thus showing a male preponderance. Klebsiella pneumoniae and Candida were the major isolates, 37.8% each. The antibiotic sensitivity showed that most of the Gram negative bacteria were highly resistant to the commonly used antibiotics like Ampicillin and Gentamicin. Gram positive bacteria showed 100% sensitivity to linezolid and Gram negative bacteria showed more than 90% sensitivity to imipenem. Conclusions: Development of sepsis in neonates is a medical emergency and generally the clinicians do not wait for microbiology report and start treatment empirically. If local microbiological databases are available with information regarding the commonly isolated organisms and their drug resistance patterns, it can help the clinicians in empirical therapy.