Background: Neonatal septicemia remains a significant healthcare challenge, particularly in resource-limited settings, with both early-onset neonatal septicemia (EONS) and late-onset neonatal septicemia (LONS) presentations contributing to morbidity and mortality. This study aimed to characterize the clinico-etiological profile and antibiotic susceptibility patterns of neonatal septicemia in a tertiary care setting in north India., Methodology: An analytical cross-sectional study was conducted from March 2021 to February 2022, encompassing neonates admitted to the Department of Pediatrics with suspected neonatal septicemia, confirmed by positive blood cultures., Results: A total of 96 neonates were included, predominantly male (71.9%). Gram-negative bacteria constituted 61.6% of isolates, and the most common organism isolated was non-lactose fermenter group (38.4%) followed by coagulase-negative staphylococci (CoNS) (33.4%). Non-lactose fermenter group bacteria were prominent in EONS cases (44.6%), while CoNS predominated in LONS cases (51.6%). Birthplace, birth weight, and perinatal score were significantly associated with both EONS and LONS. Linezolid exhibited high efficacy against gram-positive bacteria, while ciprofloxacin and meropenem demonstrated effectiveness against various gram-negative pathogens. Methicillin-resistant Staphylococcus aureus (MRSA) strains exhibited resistance to all the antibiotics used in the study except for linezolid., Conclusion: These findings underscore the importance of tailored empirical therapy guided by local epidemiological data to optimize clinical outcomes and mitigate antimicrobial resistance., Competing Interests: Human subjects: Consent was obtained or waived by all participants in this study. Institutional Ethics Committee, Indira Gandhi Medical College, Shimla issued approval HFW(MC-2)B(12)ETHICS/2021/14453 dated January 2021. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Sharma et al.)