Evaluation of antibiotics in terms of the quality of antibiotic use was carried out using the Gyssens method to evaluate the accuracy of the use of MDRO incidence antibiotics. The purpose of the study was to analyze the effect of the accuracy of antibiotic use on the incidence of MDRO and its impact on cost effectiveness analysis (CEA) and length of stay at Dr. Iskak Hospital Tulungagung. Research design in the form of quantitative research with case control analytical observational research methods with a retrospective approach. The research samples were some employees at RSUD dr. Iskak Tulunggung Regency as many as 50 case groups and 50 control groups with sampling techniques using the simple random sampling method. Data analysis using Path Analysis. The results showed that the punctuality variable of antibiotic administration had an indirect influence on the variable length of hospitalization days of -0.065, indicating that the timeliness of antibiotic administration would affect the length of hospitalization days must pass the MDRO incidence of -0.194. The variable accuracy of antibiotic administration has a direct effect on the length of days of treatment by -0.522. The variables of timeliness of antibiotic administration and the accuracy of the duration of antibiotic administration have a direct influence on the cost effective analysis of -0.191 and -0.426 without passing the MDRO event. Revise antibiogram guidelines, Increase monitoring activities for the use of antibiotics regularly, Increase teamwork of health professionals in an effort to improve the quality of antibiotic use and prevention of MDRO.