Introduction: The purpose of this study was to evaluate the sensitivity and specificity of a novel selective chromogenic agar medium (ChromID ESBL agar) that enables the detection and presumptive identification of ESBL-producing Enterobateriacae directly from clinical specimens. Materials and method: The study was conducted on 200 different clinical specimens. All samples underwent direct film examination followed by culture on MacConkey agar and the chromogenic agar media. Enterobacteriaceae isolates on MacConkey agar were screened for ESBL-production using the disc diffusion test according to the CLSI guidelines. All coloured colonies on the chromogenic agar were considered ESBL-screen positive isolates. Extended spectrum β-lactamases production was confirmed by the double disc synergy test and/or the combination disc test. Sensitivity and specificity of both screening methods were evaluated according to the results of the confirmatory tests. Results: The 200 clinical specimens yielded 221 Gram-negative bacilli isolates including 197 Enterobacteriaceae and 24 non-Enterobacteriaceae. Among Enterobacteriaceae isolates E. coli was the most common organism isolated (76%) followed by Klebsiella spp. (17%), Proteus spp (14%), Citrobacter freundii (2%) and Enterobacter cloacae (1%).Our results showed that out of 197 different Enterobacteriaceae isolates, 138 (70%) were ESBL-screen positive by the disc diffusion screening method versus 134(68%) by the chromogenic agar. The prevalence of ESBL-producing organisms was confirmed in 118 (59.8%) isolates. E. coli accounted for 78% out of them. The prevalence of faecal carriage of ESBL-producing organisms was 66.6%.The sensitivity and the negative predictive value of the disc diffusion test and the chromogenic agar were 100%. Although the chromogenic agar showed higher specificity and positive predictive value (79.7% and 88%, respectively) than the disc diffusion test (74.6% and 85.5% respectively), the difference was statistically not significant (p> 0.05).Our results revealed high concordance rate (91.8%) between the two screening methods, however the use of the chromogenic agar allowed the detection of ESBL-producers at least 24 hrs earlier than disc diffusion screening method. Also it allows direct identification of clinical isolates with no need for biochemical reaction tests. In addition the inhibitory effect of the selective chromogenic agar against the non ESBL-producing pathogens reached 32% in our study, thus allowing the reduction of unnecessary confirmatory tests. Conclusion: The chromogenic agar appears as an excellent medium for the screening and the presumptive identification of ESBL-producing Enterobacteriaecea directly from clinical samples.