Extended-spectrum beta-lactamases (ESBLs) are considered to be one of the most important antibiotic resistance mechanisms. This study reported the ESBL-producing genes in 53 randomly selected clinical bacterial isolates from the Steve Biko Academic Hospital. The presence of the bla(SHV), bla(TEM) and bla(CTX-M) genes was determined, and the overall prevalence of these genes detected in this study was 87% (46/53) in comparison with the literature; these results were higher when compared with 33% for Escherichia coli in Europe and 0.8% in Denmark for similar pathogens. These research findings indicated that it is crucial to routinely monitor the prevalence of these resistance genes.