Abstract: Antibiograms are often taken into account to define a rational selection of an empirical antimicrobial therapy for human internal fluids of treating patients with hospital-acquired infections. This study has performed a paired comparison between the antibiogram constructed with laboratory-based data and that formed with data subjected to prior clinical validation. Hence, the study was designed to determine the antibiogram of the various species of resistant pathogenic bacterium associated with hospital-acquired infection on human internal ascitic fluids. Cumulative resistance rates were estimated in parallel at the laboratory with the whole data and at the infection control department with data subjected to prior clinical validation. Results shows that no significant differences survived (p>0.05) between the percentage of isolates resistant from the infection-based system and laboratory-based system for all antimicrobial-resistant organisms studied, except methicillin resistance in Staphylococcus species. The mean difference in percentage resistance was higher from the infection-based system than the laboratory-based system for S. aureus (mean difference + 8%, p<0.001) and coagulase-negative Staphylococci (mean difference, + 9%, p<0.001). Overall, hospital antibiograms reflected susceptibility patterns among isolates associated with hospital-acquired infections. Thus, in conclusion, the Laboratory-based data underestimates the frequency of several major resistant organism in-patients with hospital-acquired infection. Previous clinical validation of the individual susceptibility reports seems to be a suitable strategy to get more reliable data.