Background: Neonatal sepsis is a clinical syndrome characterized by systemic signs of infection and accompanied by bacteremia in the first month of life. Vulnerability of neonates to infections increases with factors like low birth weight, duration of hospital stay, surgical procedures, and bacterial colonization. Method: The present study was conducted in the department of Microbiology of Maharishi Markandeshwar institute of medical science and research, Mullana, intended to characterize the clinical and microbial spectrum involved in sepsis. Result: In both suspected and culture proven sepsis cases, majority of the neonates were male and low-birth weight babies. The incidence of Gram-negative sepsis was more than gram-positive sepsis. Acinetobacter was the most common organism isolated. Conclusion: Neonatal sepsis is a substantial contributor to mortality and morbidity.