Background: Blood culture is a gold standard for accurate detection of etiological agents of infectious diseases and can assist in choice of appropriate antimicrobial therapy. So, the present study was undertaken to identify the blood culture isolates and to study the antimicrobial susceptibility pattern of blood culture isolates. Methods: Of 500 blood specimens processed from various wards and intensive care units of Mahatma Gandhi Medical College and hospitals, Jaipur, bacteria were identified and isolated. Antibiogram was performed on all positive samples. Results: 19.6% blood culture positive cases out of which 62.2% Gram positive bacteria and 37.8% Gram negative bacteria . Among Gram positive organisms, Staph aureus was found to be the predominant isolate (47.54%) followed by Coagulase negative staphylococci (CONS) (24.5%) ,Streptococcus species (19.6%) and Enterococcus species (8.19%).Methicillin resistance was found in 34.4% strains of Staph aureus.Among Gram negative organisms, most common isolates were Salmonella typhi and Acinetobacter species(32%) followed by Escherichia coli (13.5%), Klebsiella species (8%),Pseudomonas species (8%) and Citrobacter species (5%).1.02% of Candida albicans was also isolated.Conclusion: The present study showed that most of the blood culture isolates, whether Gram positive or Gram negative were multidrug resistant. The rise in antibiotic resistance in blood culture isolates emphasizes the importance of rational and judicious use of antibiotics according to the antibiotic resistance pattern of that institution.