Since the report of SHV-type enzymes in Nigeria, no survey on extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae had been performed in West African hospitals. Between May 2004 and April 2006, we analyzed 1,193 nonrepetitive Enterobacteriaceae isolates from Hoˆpital du Point G, Bamako, Mali, identified by the API 20E system. Of these, 256 produced ESBL detected by the double-disk synergy test. Enzymes were identified via PCR and sequencing: CTX-M-15 (n=157), CTX-M-14 (n=17), SHV-12 (n=17), and SHV-27 (n=1). Randomly amplified polymorphic DNA (RAPD) analysis showed great diversity of strain types, with no isolates matching E. coli O25:H4-ST 131. This study confirms that the worldwide diffusion of the CTX-M-15 enzyme has reached West Africa, probably due to plasmid spread (rather than clonal bacterial transmission), which was favored by antibiotic pressure. These findings emphasize the need for a policy of careful antibiotic use worldwide.