In this study out of 50 isolates of burns infections only 15(30%) was identified as Pseudomonas aeruginosa depending on selective media, morphological and biochemical tests then confirmed by using VITEK-2 Compact system. The pilB virulent gene was present in 15 (100%) isolates depending on PCR by using specific primers of pilB gene (826bp). The nucleotides pilB sequence has been done to nine isolate, Results displayed occurrence of some substitution gene mutations from (100-90.05%) identity with the reference sequence gene of the global strain with accession no. (pilB-879459) by alignment in NCBI and the phylogenetic tree was drew by using Geneious software. The analysis of phylogenetic tree showed five different groups of variations. The code isolates (3-pilB , 5-pilB , 9-pilB and 11-pilB) were chosen and documented in NCBI as anew Iraqi isolates and accepted under these accession numbers respectively (LC 546092, LC 546093, LC 546094 and LC 546095) of nucleotides sequence. Sequencing of pilB displayed the divergence between the Iraqi and global isolates might be due to the mutations that caused genetic variation in nucleotides content.