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Introduction: Neonatal sepsis is the most common cause of neonatal mortality. It is more common in developing countries compared with developed countries. Depending on the onset of symptoms, it can be classified into early onset sepsis within 72 h of life and late onset sepsis usually after 72 h of age. The spectrum of organisms that cause neonatal septicemia varies in different countries, and sometimes changes from one center to another within the same country. The management of these infections is complicated by the emergence of antibiotic resistance. The aim of this study was to determine the etiology, clinical characteristics and outcome of neonatal septicemia cases. Materials and Methods: One and half year study was conducted on 250 neonates admitted in the neonatal intensive care unit with clinically suspected septicemia Early onset septicemia (EOS) within 72 h of age and late onset septicemia (LOS) >72 h of age). Blood culture was performed by using BacT/Alert and detects organisms metabolic changes by fluorescence based methods which facilitates identification of microorganisms along with antibiotic susceptibility. Results: There were 210 (84%) isolates from EOS cases, while 40 (16%) were from LOS illness. In suspected as well as in culture-proven sepsis, the majority were male as compared with female. Cesarian delivary, pre term and low birth weight babies were more as compared with normal vaginal delivary, term and normal birth weight babies. Hypothermia, respiratory distress, neonatal jaundice and shock with DIC were the most common clinical presentations, and prematurity was the major risk factor in both the groups. Blood culture reports were positive in 16 (6.4%) cases of which Gram-negative bacilli and Gram positive cocci accounted for 13 (5.2%) and 2 (0.8%) respectively, in the remaining one case Candida growth was seen. Among Gram-negative bacilli, Klebsiella pnemoniae (44%) was the most common organism isolated and among Gram-positive, Enterococcus species (12%) was the commonest, followed by Burkholderia, Enterobacter, Moraxella and candida. Among Gram-positive bacteria, entrococcus species (12.5%) was the only isolate which was exhibiting MDR as well as vancomycin resistance (Strains with Van A). Strains carrying Van A genes are highly resistant to vancomycin and teicoplanin.Conclusion: Gram-negative organisms (klebsiella, Burkholderia) were the leading cause of neonatal sepsis and multidrug resistant. Therefore, an effective infection control programme, regular antibiotic susceptibility surveillance and evaluation, and the enforcement and periodic review of the antibiotic policy of the hospital as well as the encouragement of rational antibiotic use will reduce the rates of acquiring nosocomial infections and development of bacterial resistance.