Original Article

Genotypic diversity of Mycobacterium tuberculosis strains collected from immigrant patients in Mashhad, Iran using MIRU-VNTR method

Abstract

Background and Objectives: This research aimed to explore the genetic diversity and phylogenetic relationships of Mycobacterium tuberculosis (Mtb) strains, as well as to assess their drug susceptibility, specifically in strains isolated from immigrant patients attending the Referral Tuberculosis Laboratory in Mashhad.
Materials and Methods: A total of 52 sputum samples isolated from patients were examined utilizing the Mycobacterial Interspersed Repetitive-Unit Variable Number of Tandem Repeats (MIRU-VNTR). Drug-susceptibility testing against rifampin (RIF) and isoniazid (INH) was measured utilizing the proportional strategy. Thereafter, for more examination, Xpert MTB/RIF and multiplex allele-specific PCR (MAS-PCR) was performed to determine RIF and INH-resistance within the Mtb strains.
Results: Among 52 Mtb isolates, 2 (3.8%) were resistant to rifampin and one isolate was resistant to both INH and RIF and considered as multidrug-resistance (MDR) isolate. According to MIRU-VNTR, the most prominent genetic-variation patterns of these samples, were related to NEW-1 (n=18, 34.6%), followed by CAS/Delhi (n=17, 32.7%), Haarlem (n=12, 23%), Uganda I (n=2, 3.8%), S (n=1, 1.9%), Beijing (n=1, 1.9%), and unknown (n=1, 1.9%) genotypes. The statistical analysis showed that the estimated percentage of the recent TB-transmission in this study was 0.21%.
Conclusion: The result of this study indicated a great diversity of MTBC circulating among Afghan-immigrants which might be one of the reasons for the infection to become active. The relatively high percentage of resistant isolates in the studied population shows the importance of screening the immigrants especially at the entry borders and treatment and follow up of patients, to control TB-incidence in country.

1. Yang L, Zhang P, Lu S. Interpretation of World Health Organization’s Co-administration of treatment for Drug-resistant tuberculosis and Hepatitis C: 2024 update. Chin J Antitubercul 2024; 46: 874-876.
2. Williams PM, Pratt RH, Walker WL, Price SF, Stewart RJ, Feng PI. Tuberculosis - United States, 2023. MMWR Morb Mortal Wkly Rep 2024; 73: 265-270.
3. The latest status of tuberculosis in Iran in 1402. Center for Communicable Disease Management TS. behdasht.gov.ir; 2024. https://icdc.behdasht.gov.ir/TB_status/
4. Stanikzai MH, Ishaq N, Zafar MN, Baray AH, Anwary Z, Ahmad M, et al. Depression symptoms among Afghan TB patients: a multi-center study. Indian J Tuberc 2024; 71 Suppl 2: S264-S268.
5. Davuluri M, Yarlagadda VST. Novel device for enhancing tuberculosis diagnosis for faster, more accurate screening results. Int J Innov Eng Res Technol 2024; 11: 1-15.
6. Yin C, Mijiti X, Liu H, Wang Q, Cao B, Anwaierjiang A, et al. Molecular epidemiology of clinical Mycobacterium tuberculosis isolates from southern Xinjiang, China using Spoligotyping and 15-locus MIRU-VNTR typing. Infect Drug Resist 2023; 16: 1313-1326.
7. Mari S. Tipizzazione molecolare di ceppi di Mycobacterium tuberculosis complex isolati in regione Lombardia: confronto tra metodica MIRU-VNTR e Whole Genome Sequencing. 2023. https://unitesi.unipv.it/handle/20.500.14239/2/browse?type=contributorAdvisor&sort_by=ASC&order=1&rpp=20&etal=-1&authority=se22311&starts_with=A&filter_value_display=TORRI,%20STEFANIA
8. Mousavi-Sagharchi SMA, Afrazeh E, Seyyedian-Nikjeh SF, Meskini M, Doroud D, Siadat SD. New insight in molecular detection of Mycobacterium tuberculosis. AMB Express 2024; 14: 74.
9. Parsa S, Yaghoubi A, Izadi N, Sabet F, Nik LB, Derakhshan M, et al. Detection of Isoniazid and Rifampin resistance in Mycobacterium tuberculosis clinical isolates from Sputum samples by High-Resolution Melting Analysis. Curr Microbiol 2022; 79: 257.
10. Huard RC, Lazzarini LC, Butler WR, van Soolingen D, Ho JL. PCR-based method to differentiate the subspecies of the Mycobacterium tuberculosis complex on the basis of genomic deletions. J Clin Microbiol 2003; 41: 1637-1650.
11. Supply P, Allix C, Lesjean S, Cardoso-Oelemann M, Rüsch-Gerdes S, Willery E, et al. Proposal for standardization of optimized mycobacterial interspersed repetitive unit-variable-number tandem repeat typing of Mycobacterium tuberculosis. J Clin Microbiol 2006; 44: 4498-4510.
12. Allix-Béguec C, Harmsen D, Weniger T, Supply P, Niemann S. Evaluation and strategy for use of MIRU-VNTR plus, a multifunctional database for online analysis of genotyping data and phylogenetic identification of Mycobacterium tuberculosis complex isolates. J Clin Microbiol 2008; 46: 2692-2699.
13. Torkaman MR, Nasiri MJ, FaRnia P, Shahhosseiny MH, Mozafari M, Velayati AA. Estimation of recent transmission of Mycobacterium tuberculosis strains among Iranian and Afghan immigrants: A cluster-based study. J Clin Diagn Res 2014; 8: DC05-DC08.
14. Azé J, Sola C, Zhang J, Lafosse-Marin F, Yasmin M, Siddiqui R, et al. Genomics and machine learning for taxonomy consensus: the Mycobacterium tuberculosis complex paradigm. PLoS One 2015; 10(7): e0130912.
15. Weniger T, Krawczyk J, Supply P, Niemann S, Harmsen D. MIRU-VNTR plus: a web tool for polyphasic genotyping of Mycobacterium tuberculosis complex bacteria. Nucleic Acids Res 2010; 38: W326-W331.
16. Wubu B, Jemal M, Million Y, Gizachew M. Pulmonary tuberculosis and multidrug-resistant Mycobacterium tuberculosis in northwestern Ethiopia: a hospital-based cross-sectional study among presumptive pulmonary tuberculosis patients. Front Med (Lausanne) 2023; 10: 1266780.
17. Gunawan, Koraag ME, Syahnuddin M, Nursafingi A, Nurjana MA, Sumolang PPF, et al. Strain identification of Mycobacterium tuberculosis in multidrug-resistant tuberculosis (MDR-TB) patients at Undata Hospital, Palu, Central Sulawesi. Indian J Tuberc 2024; 71 Suppl 1: S15-S19.
18. Ghazvini K, Khoshbakht R, Tadayon K, Mosavari N, BahramiTaghanaki HR, Mohammadi GR, et al. Genotyping of Mycobacterium tuberculosis complex isolated from humans and animals in northeastern Iran. Sci Rep 2023; 13: 6746.
19. Naseri-Nezhad M, Asadian M, Khalifeh Gholi M, Yaseri M, Douraghi M. Mycobacterium tuberculosis genotypes in an ethnically diverse area with millions of pilgrims and thousands of immigrants. BMC Infect Dis 2023; 23: 354.
20. Riyahi Zaniani F, Moghim S, Mirhendi H, Ghasemian Safaei H, Fazeli H, Salehi M, et al. Genetic Lineages of Mycobacterium tuberculosis isolates in Isfahan, Iran. Curr Microbiol 2017; 74: 14-21.
21. Chau HTT, Nguyen PNH, Nguyen HB. Genotyping of Mycobacterium leprae strains in south central coast and central highlands of Vietnam. Iran J Microbiol 2023; 15: 201-207.
22. Khanipour S, Ebrahimzadeh N, Masoumi M, Sakhaei F, Alinezhad F, Safarpour E, et al. Haarlem 3 is the predominant genotype family in multidrug-resistant and extensively drug-resistant Mycobacterium tuberculosis in the capital of Iran: a 5-year survey. J Glob Antimicrob Resist 2016; 5: 7-10.
23. Keikha M. There is significant relationship between Beijing genotype family strains and resistance to the first-line anti-tuberculosis drugs in the Iranian population. J Clin Tuberc Other Mycobact Dis 2020; 19: 100161.
24. Richards AS, Sossen B, Emery JC, Horton KC, Heinsohn T, Frascella B, et al. Quantifying progression and regression across the spectrum of pulmonary tuberculosis: a data synthesis study. Lancet Glob Health 2023; 11(5): e684-e692.
25. Martinez L, Warren JL, Harries AD, Croda J, Espinal MA, Olarte RAL, et al. Global, regional, and national estimates of tuberculosis incidence and case detection among incarcerated individuals from 2000 to 2019: a systematic analysis. Lancet Public Health 2023; 8(7): e511-e519.
26. Fok A, Numata Y, Schulzer M, FitzGerald MJ. Risk factors for clustering of tuberculosis cases: a systematic review of population-based molecular epidemiology studies. Int J Tuberc Lung Dis 2008; 12: 480-492.
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IssueVol 17 No 3 (2025) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijm.v17i3.18817
Keywords
Mycobacterium tuberculosis Mycobacterial interspersed repetitive-unit variable number of tandem repeats Multidrug resistance

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How to Cite
1.
Jangi M, Ghazvini K, Soleimanpour S, Ghavidel M, Hashemitabar G. Genotypic diversity of Mycobacterium tuberculosis strains collected from immigrant patients in Mashhad, Iran using MIRU-VNTR method. Iran J Microbiol. 2025;17(3):358-365.