Background: Neonatal sepsis one of the most common cause for neonatal mortality and morbidity in developing countries. Group B Streptococci being the most common organism in developed countries, whereas CONS and Gram negative bacteria frequently encountered organisms in developing countries. It is advisable to have an individualised institutional protocol based on their own culture reports to reduce the antibiotic resistance.Methods:A prospective observational study was conducted from September 2016- February 2017, at the Department of Paediatrics, S.V.S. Medical College and Hospital, Mahabubnagar, Telangana, India. The study included 65 neonates admitted in our NICU. A questionnaire was filled by mothers after taking consent. Data was Collected data was analysed.Results: In present study, we found that CONS were the most commonly isolated organisms followed by Klebsiella.Conclusions:Neonatal sepsis creates a significant burden due to its impact on neonatal mortality and long-term morbidity. Following proper hand washing techniques with minimal handling in intensive care units will reduce neonatal sepsis from opportunistic infections.