Abstract

Pseudomonas aeruginosa is one of the Gram-negative bacteria widespread everywhere in the Pseudomonadaceae family and is able to survive in a wide range of environments. The aim of this study was to determine of virulent exoU genotype and phylogenetic tree of P. aeruginosa isolates using real time PCRand Sanager sequencing. 206 clinical samples were collected from wounds and burns from both sexes. Samples were cultured on MacConkey agar, Blood agar and Cetrimide agar in order to obtain the bacterial isolates depending on their phenotypic characteristics; biochemical tests as diagnosed with Api 20E; Microgen GnA + B-ID and Molecular detection of exoU mutation of Pseudomonas aeruginosa isolates by using RT-PCR and Sanger method. The results showed that 12(24%) of (50) isolates were positive for ExoU gene.The ExoU gene were used to analyze the DNA sequence of local isolates and compare them with some standard global isolates according to the ExoU gene available in the GenBank database of the NCBI database. The results showed that the isolates were (98-100%) matched between local isolates and the international standard isolates. The results of the RT-PCR of the ExoU gene were used to detect mutations related to Pseudomonas aeruginosa, and the transitions showed the frequency of 5 mutations of the ExoU gene in all isolates. In addition, there were three types of mutations such as the predicted frame mutation effect was accompanied by only a transition 3, while 4 mutations were shown silent in all isolates. In addition, 2 mutations were shown to be mismatched. Neighbor joining phylogenetic tree for ExoU sequences which indicate one cluster divided into two subgroups. Group 1 showed sister group with P. aeruginosa strain BA7823 (India) reference strain from GenBank database which was used for determining the mutations and polymorphisms in the local isolates of this study.

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