16s RNA; maximum likelihood; pseudomonas species; phylogenetic analysis; wound infection.
Purpose: Wounds infection is frequent hospital acquired infections which is caused by a wide variety of microorganism. Inappropriate treatment could be more hazardous to suffering patients and could lead to death. This manuscript is aimed to identify the existence of Pseudomonas aeruginosa from the clinical samples and determine the molecular evolution and population structure of Pseudomonas species using bioinformatics tools
Method: The specimens were swiped from the bandage of infected patients, who were admitted for the therapeutic intervention in the hospitals, localized in Delhi-NCR region. Primarly, specimens were cultured in cetrimide broth for specific growth of Pseudomonas sp. Single colony isolation on cetrimide agar plate and characterization of bacterial colony by producing fluorescence under UV light was done to confirm Pseudomonas aeruginosa (P. aeruginosa). Furthermore, phylogenetic analysis was performed on the basis of PCR and sequencing of 16s rRNA sequence of P. aeruginosa.
Results: Assay based on bacterial culture, biochemical and 16S rRNA gene analysis methods confirm the presence of P. aeruginosa in collected wound specimen. Basic local alignment search tool analysis indicates that newly sequenced 16S rRNA gene sequence was shown 99% similarity to P. aeruginosa species, analyzed using NCBI-BLAST tool. The phylogenetic analysis and nucleotide base composition studies performed using 45 sequences of 16S rRNA gene from 18 different species of Pseudomonas, including P. aeruginosa. The phylogenetic analysis was performed using Maximum Likelihood method for evolutionary relationships.
Conclusion: The biochemical and molecular characterization revealed that the clinical samples were infected with P. aeruginosa. Further, phylogenetic analysis of 16S RNA sequences revealed evolutionary conservation among different Pseudomonas species.