Background: Bloodstream infection (BSI) is a significant cause of morbidity and mortality throughout the world. Illness associated with BSI ranges from self limiting infection to life threatening sepsis that requires rapid and aggressive antimicrobial treatment. The emergence of resistant bacteria makes it a requisite to know the prevailing antibiotic susceptibility pattern of the pathogens causing bloodstream infection. Objective: To identify the bacteriological profile and antibiotic susceptibility patterns of blood culture isolates in a tertiary care hospital Material and Methods: A total of 400 patients with suspected blood stream infection from various wards in the hospital were included for this study. Blood samples were processed in microbiology laboratory and isolates were identified by standard laboratory methods and then antibiotic susceptibility test was performed by using CLSI guidelines. Results: Out of 400 blood samples, 75 (62.7%) were culture positive. Out of 75 positive cultures, 39 (52%) were gram-positive, 32 (42.7%) were gram-negative and 2 (5.3%) were Candida spp. The most predominant organism was Klebsiella species 20(26.8%), followed by Staphylococcus aureus 14 (18.67%) Most of the Gram-positive cocci (GPC) were susceptible to vancomycin and linezolid. Most of the Gram-negative bacilli (GNB) showed sensitivity to imipenem followed by amikacin. Conclusion: Increase in antibiotic resistance for BSI causing pathogens has necessitated continuous monitoring of the susceptibility of organisms towards antibiotics to prevent and spread of drug resistance.