Molecular Diagnosis of Drug-Resistant Mycobacterium tuberculosis Isolates Using IS6110 Gene in Tuberculosis Patients

Zia Sheybani, Arash; Hoseinzadeh, Shaghayegh; Habibnia, Shadi

doi:10.30699/ijmm.18.4.265

year 18, Issue 4 (July - August 2024) Iran J Med Microbiol 2024, 18(4): 265-269 | Back to browse issues page

‎ 10.30699/ijmm.18.4.265

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Zia Sheybani A, Hoseinzadeh S, Habibnia S. Molecular Diagnosis of Drug-Resistant Mycobacterium tuberculosis Isolates Using IS6110 Gene in Tuberculosis Patients. Iran J Med Microbiol 2024; 18 (4) :265-269
URL: http://ijmm.ir/article-1-2184-en.html

Molecular Diagnosis of Drug-Resistant Mycobacterium tuberculosis Isolates Using IS6110 Gene in Tuberculosis Patients

Arash Zia Sheybani¹

, Shaghayegh Hoseinzadeh²

, Shadi Habibnia³

1- Department of Microbiology, Faculty of Life Sciences, Islamic Azad University, North Tehran Branch, Tehran, Iran
2- Department of Microbiology, Faculty of Life Sciences, Islamic Azad University, Parnad Branch, Tehran, Iran
3- Department of Microbiology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran , habibnia315@gmail.com

Abstract: (629 Views)

Background and Objective: Mycobacterium tuberculosis (M. tuberculosis) is one of the most dangerous infectious diseases all over the world. Diagnosis of M. tuberculosis based on the culture and phenotypic tests takes weeks that make them inappropriate methods to evaluate a definitive diagnosis. PCR on the IS6110 gene is an accurate, quick, and inexpensive method for the M. tuberculosis detection. This study aimed to provide molecular diagnosis for the strains of M. tuberculosis resistant to the first-line drugs (rifampin and isoniazid) by IS6110 gene PCR.
Materials and Methods: Of the 3012 Mycobacterium suspected cases, 60 samples of mycobacterium drug-resistant (MDR) were examined by smear, culture, phenotypic tests, and antibiogram. The isolates were identified by IS6110 gene amplification.
Results & Conclusion: Among 60 tested samples, 54 (90%) positive samples from the M. tuberculosis group and 6 (10%) negative samples from non-tuberculosis group were reported. Out of 3012 clinical samples, 100 (3.3%) cultures were positive, 3 samples (5%) were resistant to rifampin, and the other samples (94%) were MDR. The IS6110 gene is one of the best markers for the Mycobacterium complex differentiation. This study showed that IS6110 gene PCR is a simple and accurate method for identifying drug resistance in M. tuberculosis clinical isolates. It is recommended to use molecular methods especially IS6110 gene sequencing instead of time-consuming with high variability phenotypic methods.

Keywords: Mycobacterium tuberculosis, MDR Strains, IS6110 Gene

Full-Text [PDF 486 kb] (242 Downloads) | | Full-Text (HTML) (130 Views)

Type of Study: Brief Original Article | Subject: Molecular Microbiology
Received: 2024/04/21 | Accepted: 2024/08/18 | ePublished: 2024/09/29

References

1. Cherry J, Demmler-Harrison GJ, Kaplan SL, Steinbach WJ, Hotez PJ. Feigin and Cherry's Textbook of Pediatric Infectious Diseases E-Book: 2-Volume Set. Elsevier Health Sciences; 2013.

2. Wells CD, Cegielski JP, Nelson LJ, Laserson KF, Holtz TH, Finlay A, et al. HIV infection and multidrug-resistant tuberculosis-the perfect storm. J Infect Dis. 2007;196(Supplement_1):S86-107. [DOI:10.1086/518665]

3. Agustin H, Massi MN, Djaharuddin I, Susanto AD, Islam AA, Hatta M, et al. Analysis of CD4 and CD8 expression in multidrug-resistant tuberculosis infection with diabetes mellitus: an experimental study in mice. Ann Med Surg. 2021;68:102596. [DOI:10.1016/j.amsu.2021.102596]

4. Basingnaa A, Antwi-Baffour S, Nkansah DO, Afutu E, Owusu E. Plasma levels of cytokines (IL-10, IFN-γ and TNF-α) in multidrug resistant tuberculosis and drug responsive tuberculosis patients in Ghana. Diseases. 2018;7(1):2. [DOI:10.3390/diseases7010002]

5. Kant S, Saheer S, Hassan G, Parengal J. Extra-pulmonary primary multidrug-resistant tubercular lymphadenitis in an HIV negative patient. Case Rep. 2012;2012:bcr0620114337. [DOI:10.1136/bcr.06.2011.4337]

6. Seung KJ, Keshavjee S, Rich ML. Multidrug-resistant tuberculosis and extensively drug-resistant tuberculosis. Cold Spring Harb Perspect Med. 2015;5(9):a017863. [DOI:10.1101/cshperspect.a017863]

7. Vuorinen P, Miettinen A, Vuento R, Hällström O. Direct detection of Mycobacterium tuberculosis complex in respiratory specimens by Gen-Probe amplified Mycobacterium tuberculosis direct test and Roche Amplicor Mycobacterium tuberculosis test. J Clin Microbiol. 1995;33(7):1856-9. [DOI:10.1128/jcm.33.7.1856-1859.1995]

8. Farnia P, Masjedi MR, Nasiri B, Mirsaedi M, Sorooch S, Kazeampour M, et al. Instability of IS6110 patterns in multidrug-resistant strains of Mycobacterium tuberculosis. Epidemiol Infect. 2007;135(2):346-52. [DOI:10.1017/S0950268806006790]

9. Yew WW, Leung CC. Management of multidrug‐resistant tuberculosis: update 2007. Respirology. 2008;13(1):21-46. [DOI:10.1111/j.1440-1843.2007.01180.x]

10. Tram NVQ, Bach NH, Anh NTC, Duong HH, Thanh LNX , Cappuccinelli P. Characteristics of IS6110 Sequence and Association Between Gene Mutation and Multidrug Resistance of A Multidrug-Resistant M.Tuberculosis Strain Isolated in Central Vietnam. HUE J Med Pharm. 2012;2(1):129-34.

11. Sharma SK, Kaushik G, Jha B, George N, Arora SK, Gupta D, et al. Prevalence of multidrug-resistant tuberculosis among newly diagnosed cases of sputum-positive pulmonary tuberculosis. Indian J Med Res. 2011;133(3):308-11.

12. Caminero JA. Management of multidrug-resistant tuberculosis and patients in retreatment. Eur Respir J. 2005;25(5):928-36. [DOI:10.1183/09031936.05.00103004]

13. Bhembe NL, Green E. Molecular epidemiological study of multidrug-resistant tuberculosis isolated from sputum samples in Eastern Cape, South Africa. Infect Genet Evol. 2020;80:104182. [DOI:10.1016/j.meegid.2020.104182]

14. Kabir S, Uddin MK, Chisti MJ, Fannana T, Haque ME, Uddin MR, et al. Role of PCR method using IS6110 primer in detecting Mycobacterium tuberculosis among the clinically diagnosed childhood tuberculosis patients at an urban hospital in Dhaka, Bangladesh. Int J Infect Dis. 2018;68:108-14. [DOI:10.1016/j.ijid.2018.01.015]

15. Shrestha NK, Tuohy MJ, Hall GS, Reischl U, Gordon SM, Procop GW. Detection and differentiation of Mycobacterium tuberculosis and nontuberculous mycobacterial isolates by real-time PCR. J Clin Microbiol. 2003;41(11):5121-6. [DOI:10.1128/JCM.41.11.5121-5126.2003]

16. Grange JM. The genus Mycobacterium and the Mycobacterium tuberculosis complex. In Tuberculosis: a comprehensive clinical reference. Edited by Schaaf HS, Zumla A. Philadelphia, USA: Saunders Elsevier. 2009. pp. 44-59. [DOI:10.1016/B978-1-4160-3988-4.00006-8]

17. Rohani M, Farnia P, Naderinasab M, Sadeghian A, Khajeh Karam Aldini M, Moniri R. Frequency of Beijing Genotype in Mycobacterium Tuberculosis Strains Isolated from Tuberculosis Patients in City of Mashhad. J Mazandaran Univ Med Sci. 2007;17(60):79-86.

18. Javid SN, Ghaemi EA, Amirmozaffari N, Rafiee S, Moradi A, Dadgar T. Detection of Isoniazid and Rifampin Resistant Strain of Mycobacterium Tuberculosis Isolated from patients in Golestan province (North of Iran). Med Lab J. 2009;3(1):1-8.

19. Bostanabad SZ, Salehian P, Shahraki AH. Isolation of Mycobacterium chelonae in the sputum and cervical lymph nodes of patient with metastatic breast cancer. Mycobact Dis. 2012;2(1):1000106.

20. Ruiz‐Sánchez BP, Castañeda‐Casimiro J, Cabrera‐Rivera GL, Brito‐Arriola OM, Cruz‐Zárate D, García‐Paredes VG, et al. Differential activation of innate and adaptive lymphocytes during latent or active infection with Mycobacterium tuberculosis. Microbiol Immunol. 2022;66(10):477-90. [DOI:10.1111/1348-0421.13019]

21. Kandi S, Reddy V, Nagaraja SB. Diagnosis of pulmonary and extra pulmonary tuberculosis: How best is CBNAAT when compared to conventional methods of TB detection. Pulm Res Respir Med Open J. 2017;4(2):38-41. [DOI:10.17140/PRRMOJ-4-137]

22. Alonso H, Samper S, Martín C, Otal I. Mapping IS6110 in high-copy number Mycobacterium tuberculosis strains shows specific insertion points in the Beijing genotype. BMC Genom. 2013;14:422. [DOI:10.1186/1471-2164-14-422]