Background: Sepsis related morbidity and mortality due to blood stream infections in pediatric age group is the common problem in developing countries including Ethiopia. The changing of bacteriologic profile of septicaemia and resistance to various antimicrobial agents is increasing from time to time and varied from place to place. Thus, ongoing study on the causative pathogens and their drug susceptibility pattern is paramount important in management of sepsis. Objective: To assess proportion of blood stream infection, bacterial profile and their antimicrobial susceptibility patterns among pediatric patients with suspected blood stream infection at Felege Hiwot referral Hospital. Methods: An institutional cross-sectional study was conducted on 910 blood culture and susceptibility tests was conducted in Bacteriology Laboratory of Felege Hiwot referral Hospital. The samples were collected and processed following standard microbiological techniques as part of the routine clinical management of the patient. Antibiotic susceptibility testing was done on pure culture isolates employing disc-diffusion method for the commonly used antibiotics. The data were analyzed by using SPSS version 20 and the results were summarized by using tables and graphs. Results: Out of 910 blood culture results, 444 (48.8%) were culture positive. Of all 470, (51.6%) were males and 440, (48.4%) were females and blood stream infection was highest in the age group of ≤28 days 284(64%) followed by 29 days–1 years. There was a significant association between age of patient and blood stream infection (P <0.0001) and odd of blood stream infection in age group ≤28days 4.6 fold higher AOR (95%CI=2.95-7.14). The predominant bacteria isolated from blood culture were Klebsiella spp 160(36%) followed by S. aureus 86 (19.4%) and Coagulase negative staphylococci 47(10.6%), E. coli 40(9.0%), Pseudomonas aeroginosa 33(7.43%), Enterobacter spp. 20(4.5%).The gram negative and gram positive bacteria constituted 296(66.7%) and 148(33.3%) of the culture isolates; respectively. The isolates showed high rates of resistance to most antibiotics tested. The range of resistance for Gram negative and positive bacteria were from 33%-96%, and 27.7%– 89% respectively. Ciprofloxacin, vancomycin, clindamycin, gentamycin and chloramphenicol were the most effective drugs when compared to other drugs tested against both Gram-negative and Gram positive isolates Conclusions: In the present study most of the pathogens isolated from blood culture showed high rate of resistance to most commonly used antibiotics used to treat bacterial infections. Therefore, rational use of antibiotics and infection prevention should be practiced. Keywords: Bacterial profile, antibiotic susceptibility pattern, blood stream infection, pediatric