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ICV 2023: 95.56
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Original Research Articles Volume : 15, Issue : 10, October, 2026

PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
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Editor-in-chief: Dr.M.Prakash
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Int.J.Curr.Microbiol.App.Sci.2026.15(10) : 285-295
DOI : https://doi.org/10.20546/ijcmas.2026.1510.030


Whole-Genome Sequencing and In-Silico Characterization of Mycobacterium wolinskyi from Western Rajasthan, India: Antimicrobial Resistance, Virulence Potential, and Phylogenetic Divergence

Mahima Chouhan*, Jayant Ramawat and Prabhu Prakash
Department of Microbiology, Dr Sampurnanand Medical College, Jodhpur, India
*Corresponding author
Abstract:

Mycobacterium wolinskyi is a rapidly growing nontuberculous mycobacterium (RGM) increasingly recognized as an emerging pathogen, with fewer than 30 clinical cases reported worldwide. The genomic landscape of this species remains poorly characterized, with only one draft genome sequence available in public databases. Whole-genome sequencing (WGS) of a clinical isolate CL_02207, recovered from a patient in India, was performed using Illumina paired-end sequencing. Genomic DNA was extracted using the QIAamp PowerFecal Pro DNA Kit. De novo assembly was conducted using SKESA, followed by quality assessment with QUAST. Genome annotation was performed using Prokka, and species identity was confirmed by average nucleotide identity (ANI) analysis using FastANI. Antimicrobial resistance genes were identified using AMRFinderPlus and Abricate with CARD, VFDB, and NCBI databases. Structural features including prophages, CRISPR-Cas systems, insertion sequences, and plasmids were analyzed using BLAST-based approaches. Phylogenetic analysis was performed using maximum likelihood inference with IQ-TREE. The assembled genome comprised 249 contigs totaling 6.93 Mb with an N50 of 55, 457 bp and 66.66% GC content. ANI analysis confirmed species identity (98.35% vs M. wolinskyi ATCC 700010). Prokka annotation identified 6, 788 genes including 6, 756 protein-coding sequences, 47 tRNAs, and 9 rRNAs. Six intrinsic antimicrobial resistance genes were identified (tet(V), tap, RbpA, rpoB2, vanR, lfrA), conferring resistance to tetracyclines, rifamycins, glycopeptides, and fluoroquinolones. Thirteen virulence-associated genes were detected, including regulators of iron homeostasis (ideR, mbtH), stress response (sigE, sigH, sodA), and intracellular survival (phoP, espR). No prophages, CRISPR-Cas systems, insertion sequences, or plasmids were detected. Phylogenetic analysis revealed CL_02207 as a highly divergent lineage (3.49 substitutions/site) within M. wolinskyi. This study provides the first comprehensive genomic characterization of M. wolinskyi from India, revealing intrinsic multi-drug resistance and virulence potential. The high phylogenetic divergence suggests CL_02207 represents a distinct lineage requiring further investigation. These findings contribute to the growing genomic resources for this emerging pathogen and have implications for clinical management.


Keywords: Mycobacterium wolinskyi; whole-genome sequencing; antimicrobial resistance; virulence factors; phylogenetics; nontuberculous mycobacteria; rapid-growing mycobacteria

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Mahima Chouhan, Jayant Ramawat and Prabhu Prakash. 2026. Whole-Genome Sequencing and In-Silico Characterization of Mycobacterium wolinskyi from Western Rajasthan, India: Antimicrobial Resistance, Virulence Potential, and Phylogenetic Divergence Int.J.Curr.Microbiol.App.Sci. 15(10): 285-295 doi: https://doi.org/10.20546/ijcmas.2026.1510.030
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