Pseudomonas aeruginosa possesses a variety of virulence factors that may contribute to its pathogenicity. Dissemination of resistant and highly virulent pathogens is also the main problem worldwide. Thus this study aimed to determine the frequency of oprI, Las B and toxA virulence genes among the clinical isolates of P. aeruginosa species. A total of 100 P. aeruginosa clinical isolates were collected from Hospitals and laboratories from different specimens during the period from August 2019 to November 2020. The isolates were re- identified using routine culture techniques, and multiplex PCR was applied to detect the presence of these virulence genes among them. It was found that the frequency of virulence genes were (22%) Las B, (55%) OprI, (15%) Tox A and (8%) mixed of OprI and Tox A. These genes were found with different percentage among the studied variables with the higher frequency among males, wound samples, except OprI were among 26-36 years age group and significantly among inpatients than out patients. The study concluded that all isolates carried either one or two of studied genes in different percentage while OprI represented the most predominant gene. Determination of different virulence genes of P. aeruginosa isolates suggests that they are associated with different levels of intrinsic virulence and pathogenicity.