β-lactamases exhibited by uropathogenic bacteria represent a major issue in the treatment of urinary tract infections. The resistance of β- lactamases is aworld wide medical issue. Recently, the evaluating of bacterial β- lactamases production, and identification of their drug obstruction, should be a constant cycle. The present study was performed to deduce the antibiotic resistance and the β- lactamases gene type of isolates from Oncology Center Hospital (Mansoura University). The results came out that E. coli and K. pneumoniae were the most associated strains(42.0%, and35.0%, respectively). Antimicrobial sensitivity test displayed that K. pneumoniae was resistance to Ciprofloxacin and Levofloxacin, Cefotriaxon and Trimethoprim- Sulpham by 37.8%, 39.1%, 39.3%, and 33.8%, respectively. E. coli was resistant to Ciprofloxacin and Levofloxacin, Cefotriaxon and Trimethoprim- Sulpham by 41.8%,41.8%,45.4%, and46.7%, respectively. The detection of TEM gene using Polymerase Chain Reaction showed that it was introduce by 70% in K. pneumoniae and 30% in E.coli of isolates. It could be concluded that β- lactamases production among uropathogenic bacteria is present at a high rate among urinary tract Egyptian cases in Oncology Center Hospital,. Mansoura University.