Background Irrational use of antibiotics could increase the incidence of infection caused by the extended-spectrum beta-lactamase (ESBL) producing Enterobacteriaceae. Prevalence of ESBL-producing Enterobacteriaceae varies among hospitals, and its resistance could complicate the treatment, extend the hospital length of stay, and increase the mortality. The aim of this study is to determine the prevalence of ESBL-producing Enterobacteriaceae and its association with antibiotic use. Methods This study was a cross sectional study, involving 111 secondary data derived from ESBL resistance test of culture examinations which used the standardized method according to The Clinical and Laboratory Standards Institute (CLSI) and patients medical records in Central ICU RSCM in 2011. Results Laboratory test results showed that 8 from 111 samples (7,2%) were infected with Enterobacteriaceae producing ESBL. Data were analyzed using chi-square test, p=0,05. Statistical analysis results were RP>1 with the value of significance p=1.000 and 95% CI 1.039; 1.179. Conclusion It is concluded that there is no association between antibiotic use and ESBL-producing Enterobacteriaceae infection in Central ICU RSCM in 2011.