With the worldwide spread of antibiotic resistance, delivering antibiotic susceptibility test (AST) results in a timely manner represents a major challenge. In cases of sepsis, rapid AST may facilitate early optimization of empiric antibiotic therapy. Disc diffusion is a well-standardized AST method, however 16 to 24 h are required to achieve an overall AST profile according to antimicrobial societies. In this prospective pilot study, we evaluated the performance of Mueller-Hinton-Rapid-SIR (MHR-SIR) agar after 6–8 h of incubation in comparison with standard MH agar after 16 h of incubation directly on positive blood cultures caused by Enterobacteriaceae and Staphylococcus aureus from routine clinical microbiology. A total of 133 positive blood samples including 110 Enterobacteriaceae (83%) and 23 Staphylococcus aureus (17%) were tested in parallel by two direct AST methods, each using EUCAST breakpoints. For each combination bacterium and antibiotic, we compared the categorical agreement and the correlation between the diameters obtained by MHR-SIR and by standard MH. Our results showed 97.7% categorical agreement for Enterobacteriaceae, with 1.4% minor errors, 0.4% major errors and 0.5% very major errors. For S. aureus, we observed 97.8% categorical agreement, 1.9% minor errors, 0.3% major errors and no very major errors. Our results showed excellent categorical agreement and correlations between diameters for MHR-SIR and standard MH methods. MHRSIR can predict the result of overall AST profile within 6–8 h with reliable results. AST is obtained on the same day the blood culture becomes positive, with a very moderate cost. •First evaluation of Mueller-Hinton Rapid-SIR agar (MHR-SIR) directly from blood cultures•Time gain of 8 h compared to standard methods with a read of a non-expert microbiologist•This study showed excellent categorical agreement and correlations between diameters for MHR-SIR and standard MH methods.•Allow early antibiotic appropriateness in 6–8 h after positivity of blood culture and this accelerate prescription of targeted antimicrobial treatment thereby potentially improving the patients' clinical outcome. This appropriateness of empirical antimicrobial therapy allows a de-escalation of broad-spectrum antimicrobial agents.