Pseudomonas aeruginosa is identified as an opportunistic pathogen since it predominantly creates nosocomial infections in immune compromised people; total of 20 samples of P. aeruginosa obtained from a variety of locations in Baghdad hospitals isolated from: burns, wounds and sputum collected from October 2021 to the April 2022, isolates cultured by Using culture conditions, biochemical assays, and the VITEK-2 compact system. For all bacterial isolates, Biofilm production was detected using the micro titration plate technique, with the majority of strains (50%) producing moderate biofilms, 35% producing strong biofilms, and 15% producing weak biofilms. When it comes to virulence genes, In 20 isolates, three genes were identified, the algD gene was found in 17 isolates (85%), pelA was found in 17 isolates (85%), and pslD was found in 15 isolates (75%). The genes algD, pslD, and pelA were chosen as virulence factors, the genotypic algD/pslD/pelA – pattern, which is responsible for the phenotypic pattern of biofilm development. There were statistically significant variations in genotypic pattern predominance across biofilm forming isolates from various sources, Using Chi-square analysis, a very significant relationship between biofilm forming capability and genetic pattern (p value 0.764) was discovered.