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Background: Pseudomonas aeruginosa is a Gram-negative bacterium, and it is a pathogen of opportunistic diseases that affects human with immunodeficiency or breaches of normal bodily defenses. Aims: Isolation of P. aeruginosa bacteria from administration samples from patients with UTI, diagnosing this bacterium using enriched and selective culture media in addition to the biochemical assays. It also aims to extract bacterial DNA from isolates to investigate the lasB and plcH genes responsible for some virulence factors in P. aeruginosa, using the Polymerase chain reaction (PCR) technique. Materials and methods: The current study included the collection of 477 samples of patients suffering from Urinary tract infections (UTIs) of both sexes, whose ages ranged between (19-65) years, for the period from September 1, 2020 to December 31, 2020, from several hospitals in the city of Baghdad. Results: After the final diagnosis of the samples, 50 (10.48%) isolates of P. aeruginosa were obtained. The percentage of infection with this bacteria was higher in males than in females, 26 (52%) and 24 (48%), respectively. The highest rate of bacterial infection was in the age group (51-60) years, at 38%. The isolates were tested for antibiotic sensitivity using the Kirby-Bauer method, and the highest antibiotic resistance was Ceftazidime (98%), Aztreonam (96%), Amikacin (76%) and Tobramycin (74%). DNA was extracted from all bacterial isolates, and the polymerase chain reaction (PCR) was performed using specialized primers to investigate the lasB and plcH genes. The results showed that 24 (48%) isolates contain the lasB gene, and the resulting bundles were found to have a molecular weight (284 bp). The results also revealed that 24 (48%) isolates contain plcH gene with a molecular weight of the bundles (608 bp). Conclusion: it was found in the current study that the bacterial isolates that contained the lasB gene are the same as that containing the plcH gene, so the possibility of these isolates for pathogenicity is higher than if they contained only one of the two genes.