Molecular Diagnosis Of Tuberculosis and Non-Tuberculous Mycobacteria: Evaluating Dual PCR Targeting IS6110 and rpoB Gene
Keywords:
Dual PCR, IS6110, rpoB, MTB, NTMAbstract
A sum of 100 clinical specimens was examined, showing that sputum had the highest positivity rates for MTB, with 13 of 34 samples testing positive for rpoB and 9 for IS6110. Significantly, differences among these markers were noted, indicating possible genetic variations or co-infections. Pus samples (n=26) demonstrated a low detection rate, with merely 6 samples testing positive for IS6110, while fistula samples (n=18) exhibited an equal distribution of rpoB and IS6110 positivity (7 cases each). Menstrual blood samples (n=9) presented distinct diagnostic challenges, showing rpoB positivity in 2 instances and IS6110 in 3 instances. In total, MTB was detected in 35 instances (70%), whereas NTM was verified in 15 instances (30%). These results highlight the importance of multi-gene molecular diagnostics for accurate distinction between MTB and NTM, decreasing the chances of misdiagnosis. The research shows that dual PCR is a strong and dependable diagnostic method, potentially enhancing TB case identification and focused treatment approaches. Additional validation involving varied populations and incorporation with next-generation sequencing is advised to improve diagnostic precision and public health strategies.
Downloads
Metrics
References
World Health Organization (WHO). (2022). Global Tuberculosis Report 2022. WHO Press.
Daley, C. L., Iaccarino, J. M., Lange, C., Cambau, E., Wallace, R. J., Andrejak, C., ... & Griffith, D. E. (2020). Treatment of nontuberculous mycobacterial pulmonary disease: an official ATS/ERS/ESCMID/IDSA guideline. European Respiratory Journal, 56(1), 2000535.
Forbes, B. A., Hall, G. S., Miller, M. B., Novak, S. M., Rowlinson, M. C., Salfinger, M., & Somoskövi, Á. (2018). Practice guidelines for clinical microbiology laboratories: Mycobacteria. Clinical Microbiology Reviews, 31(2), e00038-17.
van Soolingen, D., Hermans, P. W., de Haas, P. E., Soll, D. R., & van Embden, J. D. (1993). Occurrence and stability of insertion sequences in Mycobacterium tuberculosis complex strains: evaluation of an insertion sequence-dependent DNA polymorphism as a tool in the epidemiology of tuberculosis. Journal of Clinical Microbiology, 31(8), 1961-1965.
Kim, B. J., Math, R. K., Jeon, C. O., Yu, H. K., Park, Y. G., Kook, Y. H., & Kim, B. J. (2013). A PCR-based method for discrimination of Mycobacterium tuberculosis complex and nontuberculous mycobacteria using rpoB gene-derived primers. Diagnostic Microbiology and Infectious Disease, 75(2), 175-176.
Huang, W. C., Yu, M. C., Chen, H. Y., Hsu, H. L., & Jou, R. (2019). Evaluation of a multiplex real-time PCR for identification of Mycobacterium tuberculosis complex and nontuberculous mycobacteria. Journal of Microbiological Methods, 158, 1-7.
Singh, N., Jain, S., Goel, A., Bhargava, R., & Verma, R. (2021). Application of dual PCR for rapid identification of mycobacterial infections: An alternative to conventional methods. Indian Journal of Medical Microbiology, 39(1), 72-78.
Ryu, Y. J., Koh, W. J., Daley, C. L., & Kim, S. Y. (2022). Diagnostic advances for pulmonary tuberculosis and nontuberculous mycobacterial lung disease. Expert Review of Molecular Diagnostics, 22(4), 373-389.
Brown-Elliott, B. A., Philley, J. V., & Wallace, R. J. (2019). Current and emerging diagnostic approaches for nontuberculous mycobacterial infections. Clinical Microbiology Reviews, 32(2), e00010-19.
Koh, W. J., Stout, J. E., & Yew, W. W. (2020). Advances in the diagnosis and treatment of pulmonary disease due to Mycobacterium avium complex. Respirology, 25(10), 1022-1035.
Parekh, K. M., Inamdar, V., Jog, A., and Kar, A. (2006). A comparative study of the diagnosis of pulmonary tuberculosis using conventional tools and polymerase chain reaction. Indian Journal of Tuberculosis, 53, 69-76.
Tang, Y. W., Meng, S., Li, H., Stratton, C. W., Koyamatsu, T., and Zheng, X. (2004). PCR enhances acid-fast bacillus stain-based rapid detection of Mycobacterium tuberculosis . Journal of Clinical Microbiology, 42(4), 1849-1850. https://doi.org/10.1128/JCM.42.4.1849-1850.2004.
Kim, Bum-Joon, Seong-Karp Hong, Keun-Hwa Lee, Yeo-Jun Yun, Eui-Chong Kim, Young-Gil Park, Gil-Han Bai, and Yoon-Hoh Kook. Journal of clinical microbiology 42, no. 3 (2004): 1308-1312.
Zhao, Y, (2019). Molecular detection of non-tuberculous mycobacteria using PCR and sequencing methods. Diagnostic Microbiology and Infectious Disease, 95, 137–142. https://doi.org/10.1016/j.diagmicrobio.2019.03.003
Goletti, D.,(2022). Current and future diagnostic approaches for tuberculosis: An overview of advances and challenges. Expert Review of Molecular Diagnostics, 22, 115–130. https://doi.org/10.1080/14737159.2022.2040468
Boehme, C. C., Nabeta, P., Hillemann, D., Nicol, M. P., Shenai, S., Krapp, F., ... & Perkins, M. D. (2011). Rapid molecular detection of tuberculosis and rifampin resistance. New England Journal of Medicine, 363(11), 1005-1015. https://doi.org/10.1056/NEJMoa0907847
Mokrousov, I., Narvskaya, O., Otten, T., Limeschenko, E., Steklova, L., & Vyazovaya, A. (2016). High prevalence of rifampin-resistant M. tuberculosis strains in a TB hospital in St. Petersburg, Russia: a retrospective study. Journal of Clinical Microbiology, 40(11), 4015-4018.
Telenti, A., Imboden, P., Marchesi, F., Lowrie, D., Cole, S., Colston, M. J., ... & Bodmer, T. (1993). Detection of rifampicin-resistance mutations in Mycobacterium tuberculosis. The Lancet, 341(8846), 647-650. https://doi.org/10.1016/0140-6736(93)90444-W
Koh, W. J., Chang, B., Jeong, B. H., Jeon, K., Kim, S. Y., & Kim, C. K. (2020). Clinical significance and treatment outcomes of pulmonary disease caused by Mycobacterium abscessus and Mycobacterium massiliense. Clinical Infectious Diseases, 70(5), 754-762. https://doi.org/10.1093/cid/ciz285
Downloads
Published
How to Cite
Issue
Section
License

This work is licensed under a Creative Commons Attribution 4.0 International License.
You are free to:
- Share — copy and redistribute the material in any medium or format
- Adapt — remix, transform, and build upon the material for any purpose, even commercially.
Terms:
- Attribution — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use.
- No additional restrictions — You may not apply legal terms or technological measures that legally restrict others from doing anything the license permits.