Abstract Pseudomonas aeruginosa is a potent nosocomial pathogen, causing several infections, mostly urinary tract infections (UTIs). The present study is thus aimed to detect the susceptibility pattern of Pseudomonas aeruginosa to antibiotics from urine specimens, and virulence factor production such as (biofilm, and pyocyanin) which are regulated by quorum sensing. The aims were extended to detect the Inhibition of Biofilm formation and pyocyanin production by using vitamin C, salicylic acid and Multi sera. A total of 245 samples were collected from the patients. The samples were subjected to inoculation, isolation and identification of Pseudomonas aeruginosa which were 8 (4%) from total isolates, by standard microbiological procedures. Confirmation of isolates was done by polymerase chain reaction targeting 16srRNA. Antimicrobial sensitivity testing was done using the modified Kirby-Bauer method of the disc diffusion test, with high resistance rates against ceftriaxone (100%) while h