Background- Blood stream infection is a very important cause of morbidity and mortality worldwide, especially in resource limited countries. It ranges from transient bacteremia to life-threatening septic shock. Blood culture is a gold standard method of diagnosis of infectious agents present in the blood.Method- This study was conducted From August 2021 to October 2021 at Ruxmaniben Deepchand Gardi medical and Ujjain Charitable trust Hospital, Ujjain (M.P.). All received blood samples were processed, isolates were identified, and antibiotic susceptibility testing was performed using the Vitek 2 Compact (Biomerieux) system in the Microbiology laboratory.Results- A total of 74(17%) pathogens were isolated from 432 blood samples. Gram-positive bacteria 47(64%) were predominant organisms obtained, followed by gram-negative bacteria 25(34%) and non-albicans candida species 2(2.7%). The predominant pathogens were Staphylococcus aureus 40(54%), Klebsiella pneumoniae and Pseudomonas aeruginosa 6(24%) each. Among Staphylococcus aureus isolates 36 (90%) were methicillin-resistant Staphylococcus aureus (MRSA). And among gram-negative bacteria, all isolates of K. pneumoniae were extensively drug-resistant. Majority of drug-resistant organisms were isolated from neonatal intensive care unit (%).Conclusion- Successful treatment of bloodstream infection depends on early diagnosis and appropriate use of antimicrobial agents.