Case Control Study
Copyright ©The Author(s) 2025. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Exp Med. Sep 20, 2025; 15(3): 105208
Published online Sep 20, 2025. doi: 10.5493/wjem.v15.i3.105208
Evaluation of urinary extracellular vesicles and microRNAs to diagnose urogenital tuberculosis
Parijat Das, Dharmendra K Chaudhary, Richa Mishra, Swasti Tiwari
Parijat Das, Dharmendra K Chaudhary, Department of Molecular Medicine and Biotechnology, Sanjay Gandhi Post-Graduate Institute of Medical Sciences, Lucknow 226014, Uttar Pradesh, India
Richa Mishra, Department of Microbiology, Sanjay Gandhi Postgrad Institute Medical Sciences, Lucknow 226014, Uttar Pradesh, India
Swasti Tiwari, Department of Molecular Medicine and Biotechnology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow 226014, Uttar Pradesh, India
Author contributions: Das P carried out data extraction; Das P and Chaudhary DK performed data curation; Das P and Mishra R contributed to formal analysis; Tiwari S and Mishra R contributed to methodology, reviews and editing; Tiwari S carried out conceptualization and fund acquisition; Das P, Chaudhary DK, and Tiwari S wrote the article; All authors contributed to the interpretation of the study and approved the final version to be published.
Supported by the Indian Council of Medical Research, No. Coord/7 (1)/CARE-KD/18-NCD-II.
Institutional review board statement: This study was approved by the Institutional Ethics Committee of Sanjay Gandhi Postgraduate Institute of Medical Sciences, approval No. PGI/BE/25/2023.
Informed consent statement: Subjects have signed informed consent.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
STROBE statement: The authors have read the STROBE Statement-checklist of items, and the manuscript was prepared and revised according to the STROBE Statement-checklist of items.
Data sharing statement: The data supporting the findings of this study are available from the corresponding author upon reasonable request.
Open Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Swasti Tiwari, PhD, Professor, Department of Molecular Medicine and Biotechnology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Raebareli Road, Mawaiya, Lucknow 226014, Uttar Pradesh, India. tiwari.pgi@gmail.com
Received: January 15, 2025
Revised: April 22, 2025
Accepted: June 20, 2025
Published online: September 20, 2025
Processing time: 210 Days and 0.3 Hours
Abstract
BACKGROUND

India has the highest tuberculosis (TB) burden in the world. Of the estimated annual 10 million TB cases, features of extra pulmonary TB are evident in up to 45%. Urogenital TB (UGTB) accounts for approximately 20% of those cases. The lack of non-sputum based diagnostic tools continue to hinder efforts to reduce the burden of UGTB. MicroRNAs (miRNAs) play a crucial role in biological pathways and can be used as a potential biomarker for TB. We evaluated urinary extracellular vesicles (uEVs) as non-invasive source to explore miRNAs with biomarker potential for UGTB.

AIM

To evaluate the potential of miRNA-155-5p, miRNA-26a-5p and miRNA-29a-3p in uEVs to diagnose UGTB in adults.

METHODS

uEV characterization was done using nanoparticle tracking analysis and flow cytometry. Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) for urinary uEV-miRNAs were carried out in samples from patients with suspected UGTB, or Urinary tract infections [UTI, disease controls (DC)] and healthy controls (HCs) (n = 20/group). U6 was used to normalize the qRT-PCR data. Receivers operating characteristic curves was used to calculate the diagnostic accuracy of uEV-miRNAs to differentiate UGTB from controls (DC and HCs).

RESULTS

uEVs from UGTB or UTI patients had higher mean size, and also lower proportion of CD63 positive vesicles as compared to HC’s uEVs. Between UTI and UGTB, the mean size of uEVs was significantly higher in UTI cases. qRT-PCR analysis revealed a significantly lower abundance of miRNA-155-5p and miRNA-26a-5p in uEVs from UGTB relative to UTI (P value = 0.004) and HC (P value = 0.009) respectively n = 20/group). While, miRNA-29a-3p was higher in abundance in both UGTB and HCs’ uEV, relative to uEVs from UTI cases (P values = 0.004 and 0.002 respectively, n = 20/group). Moreover, miRNA-155-5p [area under curve (AUC) = 0.88, P ≤ 0.0001] and miRNA-29a-3p (AUC = 0.76, P value = 0.005) had optimal diagnostic accuracy to differentiate UGTB from DC (n = 20/groups) with a likelihood ratio of 5.2 and 4.3, respectively through receivers operating characteristic curve. While, miRNA-155-5p (AUC = 0.68, P value = 0.05) and miRNA-26a-5p (AUC = 0.78, P value = 0.002) had optimal diagnostic accuracy to differentiate UGTB from HCs with a likelihood ratio of > 2.

CONCLUSION

The differential expression of uEV-miRNAs, miRNA-155-5p and miRNA-29a-3p in UTGB and UTI cases hold promise in the specific diagnosis of UGTB. Further studies in large cohort are, however, needed to confirm the diagnostic accuracy of these uEV-miRNAs.

Keywords: Tuberculosis; Urine; Extracellular vesicles; MicroRNA; Biomarker

Core Tip: Discovery of new biomarkers from easily attainable bodily fluids is essential for controlling Urogenital tuberculosis (TB), a significant health concern. Urinary extracellular vesicle microRNAs (miRNAs) are exciting as biomarkers given their stability and non-invasive nature. miRNA-155, miRNA-26 and miRNA-29 play a crucial role in mycobacterium TB pathogenesis via regulation of posttranscriptional gene expression and subsequently apoptosis, cellular proliferation, differentiation and several other biological functions governing various facets of innate and acquired immunity. Thus, there is scope for developing miRNA studies using in vitro and in vivo models in TB which would likely lead to new technologies in diagnosis and treatment.