In Mali, urinary infection is one of the most common bacterial diseases in outpatients. Treatment of these diseases has become challenging due to the emergence of pathogens with increasing resistance to available antimicrobial agents. The aim of this study is to determine the prevalence and evaluate the antibiotic resistance profile of the most frequently isolated enterobacteria strains involved in urinary infections in Bamako, Mali. In this study, 1050 patients with clinical symptoms of urinary infection, referred to "Institut National de Recherche en Santé Publique (INRSP)″ of Bamako, Mali, were evaluated between December 2010 and January 2011. The isolated bacteria were identified by biochemical tests. Disk diffusion method was applied to determine the antibiotic sensitivity of bacterial agents. Our results showed that 231 (22%) patients out of 1050 were shown to be urine culture positive (70.6% females and 29.4% males). The most isolated entero-bacterium was E. coli with frequency rate of 62.3%. The other enterobacteria were Klebsiella pneumoniae (19.5%), Enterobacter cloacea (6.1%), klebsiella oxycota (4.3%) and Proteusmirabilis (2.2%). All the enterobacteria isolated from urine specimens were sensitive to amikacin and colistin (92.9%-100%). The amikacin was more effective against all the uropathogen enterobacteria isolated in present study. According to the present survey, E. coli isolates were the predominant pathogens in urinary infections in the district of Bamako and were more prevalent in women than men. In the present study, the frequency of urinary pathogens was comparable to that reported in previous studies, showing an increasing resistance pattern to commonly prescribed antibiotics.