Original Research

Antibiotic Susceptibility Profiling, Molecular Characterization, and16S rRNA-Based Phylogenetic Analysis of Pseudomonas Aeruginosa Isolates from Musculoskeletal Infections

Wajeeha Batool1
Institute of Molecular Biology and Biotechnology (IMBB), The University of Lahore, Pakistan
Maria Muddassir2
M Islam Medical College, Gujranwala, Pakistan
Muhammad Junaid Iqbal3
Sahiwal Medical College, Sahiwal, Pakistan
Abdul Ghaffar3
Sahiwal Medical College, Sahiwal, Pakistan
Muhammad Qasim Ali4
Gulab Devi Educational Complex, Lahore, Pakistan
Syed Zeeshan Haider Naqvi1
Institute of Molecular Biology and Biotechnology (IMBB), The University of Lahore, Pakistan

✉ Correspondence: Syed Zeeshan Haider Naqvi Email: zeeshan.haider@imbb.uol.edu.pk

DOI https://doi.org/10.70302/jpsim.v6i2.2535
Volume: 6, Issue 2 Pages: 185–190 Published: 05 Jun 2025
Received: 09 Jan 2025 Revised: 25 Feb 2025 Accepted: 28 Apr 2025 Published Online: 05 Jun 2025

Abstract

Objective

The primary goal of this research was to describe antibiotic resistance and identification of virulence genes in Pseudomonas aeruginosa.

Methods

The study was cross-sectional, featuring a randomized selection of samples. Conducted from January to December 2023, the study encompassed a sample size of 320 including blood, pus, and wound swabs [Male:169(52.8%), Female 151(47.1%)]. Age-based categories were ascertained, G1:13-20 years, G2:21-40 years, G3:41-60 years, G4:61-80 years. Identification tests for Pseudomonas aeruginosa included API20NE (bioMérieux®, France). Antimicrobial susceptibility testing was following the CLSI,2020 for the Disc Diffusion Test.DNA extraction and purification were carried out through a Genome Jet DNA Purification kit and 16S rRNA primers for the identification of resistance genes exoA and oprL. Phylogenetic and taxonomic identification was conducted via ribosomal RNA sequencing of PCR-amplified products(BIO-RAD T100TM Thermocycler) and sequencing with an automated sequencer (Illumina MiniSeqTM). Phylogenetic tree was constructed by MEGA11.p-value<0.05 was statistically significant.

Results

Prevalence of Pseudomonas aeruginosa was 22%. Positivity was recorded from pus (152/320:47.5%),blood(124/320-38.75%) and wound swabs(44/320-1 3.75%).G4 showed maximum positive isolates(91.25%).Maximum resistance was exhibited against Meropenem(76%) and Imipenem(70%).PCR identified the presence of resistance genes,exoA gene(125 bp), and oprL(105 bp). The phylogenetic tree was constructed through ribosomal RNA.The gene bank accession number for 16S rRNA gene of P. aeruginosa is PQ269824.

Conclusion

Antimicrobial susceptibility testing helps improve the treatment alternatives for stubborn strains. The presence of resistance genes highlights how research at the molecular level enhances treatment approaches for managing multi-drug resistant pathogens.

Keywords
Pseudomonas aeruginosa antibiotic susceptibility musculoskeletal infections phylogenetic tree.

How to Cite This Article

Batool W, Muddassir M, Iqbal MJ, Ghaffar A, Ali MQ, Naqvi SZH. Antibiotic Susceptibility Profiling, Molecular Characterization, and16S rRNA-Based Phylogenetic Analysis of Pseudomonas Aeruginosa Isolates from Musculoskeletal Infections. J Pak Soc Intern Med. 2025;6(2):185–190. doi:10.70302/jpsim.v6i2.2535

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Conflict of Interest

All authors declare no competing interests.

Disclosed in accordance with ICMJE and COPE guidelines.

Funding

No specific funding was received for this research.

Funder information follows the Crossref Funder Registry standard.

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CC BY CC BY 4.0 This article is published under the Creative Commons Attribution 4.0 International License. May be shared and adapted with attribution. © 2025 Journal of Pakistan Society of Internal Medicine.
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