Molecular identification and virulence analysis of pseudomonas aeruginosa isolated from burns, wounds and uti patients in Babylon, Iraq

Pseudomonas aeruginosa, a Gram-negative bacterium, is of great concern due to its resistance to antibiotics. Researchers are studying its innate mechanisms, such as efflux pumps and antibiotic targets, and resistance acquired through gene transfer. This highlights the need for strategies in microbial ecology and disease management. In our study, we investigated P. aeruginosa in Babylon City, Iraq, focusing on its diverse genetic structure and pathogenicity. The research aims to understand the epidemiology of P. aeruginosa infections and develop targeted treatment strategies. The samples consisted of 100 participants, divided into burns, wounds and urinary tract infections, and a control group of 50 healthy individuals, totalling 150 individuals. VITEK-2 system was used to determine the MIC. Biofilm production rate was estimated by microtiter plate method. DNA was then extracted, ExoA and FliC genes were amplified by PCR and the amplified product was sequenced. The sequences obtained were then analysed using NCBI Blast tools to detect various nucleotides. The results of Pseudomonas aeruginosa culture showed typical results of bacterial culture. Analysis of the data shows that the highest percentage of P. aeruginosa isolates (30%) was found in the 17-26 age group. 28% was found in the 27-36 age group, 24% in the 37-45 age group and the lowest percentage was found in the 46-51 age group with 18%. The chi square value was 1.680 and the corresponding p-value was 0.641. In the distribution of the samples according to the infection site, the highest infection site was wound isolates with 42%, followed by burns with 36% and urinary tract samples with 22% (chi-square value 3.160 and p-value 0.206). The distribution of positive samples was evenly distributed between both sexes (male and female), with a P-value equal to 0.631. The distribution according to blood groups shows that 50 positive isolates were distributed among eight blood groups; O+ (16%), O (12%) A+ (16%) A (10%) B+ (14%) B (12%) AB+ (12%) and AB (8%). The chi-square value is 2.160 and the p-value is as high as 0.950. The MIC results obtained with the VITEK-2 system revealed that there was resistance among the strains tested, with the lowest sensitivity to Gentamicin (GEN) at 78% and the lowest sensitivity to Aztreonam (ATM) at 0%. On the other hand, the highest resistance was recorded against Meropenem (MRP) with 96%, while iv Gentamicin (GEN) showed 22% resistance. This comparison was accompanied by a significant P-Value equal to 0.004. The results of amplification followed by sequencing showed different variations in the ExoA gene compared to the reference sequence CP075787.1 in the NCBI database. The isolate showing the highest variation rate also showed the highest biofilm production rate. Sequence analysis of the Flic gene also showed different variations when compared to the reference sequence with reference ID (L81147.1.). The samples with the highest variation rate showed high MIC levels. From the results of this study, it can be concluded that Pseudomonas aeruginosa infection is common in young adults. Gentamicin is effective in the treatment of most P. aeruginosa isolates due to high susceptibility rates. Genetic variations are positively correlated with biofilm production. Isolates with higher ExoA and FliC gene variations produced more biofilms. There is a positive correlation between genetic variations of ExoA and FliC genes and MIC. High antibiotic resistance is associated with gene variations.