Extended spectrum β-lactamase (ESBL) and carbapenemase-producing Enterobacteriaceae (CPE) have been increasingly reported worldwide. The objective of this study is to determine the prevalence of these multidrug-resistant strains in a major university teaching hospital in Dakar, Senegal. A total of 1205 Enterobacteriaceae stains were tested for ESBL and carbapenemase production. Antibiotics susceptibility test was performed with disk diffusion method. ESBL was detected using a double-disk synergy method. Carbapenemase production was detected with ertapenem 10 µg disk charge. The overall prevalence of ESBL-and carbapenemase-producing Enterobacteriaceae was 26.2 (316/1205) and 5.1% (62/1205), respectively. Interestingly, 3.8% of these pathogens were both ESBL-carbapenemase producers. Among the Enterobacteriaceae ESBL positive, Escherichia coli (45.2%, 143/316), Klebsiella pneumoniae (26.3%, 83/316), Enterobacter cloacae (12.7%, 40/316), and Proteus vulgaris (9.2%, 29/316) were the most prevalent. These strains were mainly isolated from urine (56.6%) and pus (22.7%) specimen. The most prevalent CPEs were E. coli (45.2%, 28/62), K. pneumoniae (27.4%, 17/62), and E. cloacae (16.1%, 10/62), particularly isolated from urine (58%) and pus (19.3%). The majority of these MDR strains were isolated from patients hospitalized in urology (32.4%), surgery (27.7%), internal medicine (18.5%), and intensive care units (10%). ESBL-producing Enterobacteriaceae remain highly susceptible to fosfomycin (94.1%), amikacin (92.5%) and ertapenem (88.6%), while carbapenemase producers were fully susceptible to amikacin (100%), and to a lesser extent, fosfomycin (66.7%) and colistin (60%). This study revealed increasing prevalence of ESBL-and carbapenemase-producing Enterobacteriaceae with limited therapeutic options, suggesting a need for continuous multi-drug resistant (MDR) surveillance patterns particularly in hospital settings.