Investigation of Drug Resistant Mycobacterium tuberculosis Strains with Molecular Methods
Yıl 2025,
Cilt: 51 Sayı: 2, 291 - 294, 28.08.2025
Ayşe Melda Payaslıoğlu
,
Sezcan Sağlam
,
Seçil Ak Aksoy
,
Cüneyt Özakın
Öz
The aim of this study was to investigate mutations in the rpoB and embB genes using molecular methods in Mycobacterium tuberculosis isolates phenotypically resistant to rifampicin and/or ethambutol. Drug susceptibility testing of 110 isolates collected in Turkey between 2005 and 2014 was performed using the modified agar proportion method. Molecular analysis was conducted on 28 selected isolates, including 7 rifampicin-resistant strains. Mutations were detected within the Rifampicin Resistance-Determining Region (RRDR) between codons 507–533 of the rpoB gene. The most common mutation was Ser531Leu (TCG→TTG), found in 10 strains; one strain had a His526Asn (CAC→AAC) mutation. These mutations were considered important molecular markers of rifampicin resistance. Rapid molecular detection of these mutations is important for early and effective tuberculosis treatment planning.
Kaynakça
-
1. Gagneux S. Ecology and evolution of Mycobacterium tuberculosis. Nat Rev Microbiol. 2018;16(4):202–213. doi:10.1038/nrmicro.2018.8
-
2. Pai M, Behr MA, Dowdy D, Dheda K, Divangahi M, Boehme CC, et al. Tuberculosis. Nat Rev Dis Primers. 2016;2:16076. doi:10.1038/nrdp.2016.76
-
3. World Health Organization. Global Tuberculosis Report 2023. 2023. [Accessed: June 23, 2024]
-
4. Dong B, He Z, Li Y, Xu X, Wang C, Zeng J. Improved conventional and new approaches in the diagnosis of tuberculosis. Front Microbiol. 2022;13:924410.
-
5. Boehme CC, Nabeta P, Hillemann D, Nicol MP, Shenai S, Krapp F, et al. Rapid molecular detection of tuberculosis and rifampin resistance. N Engl J Med. 2010;363(11):1005–1015. doi:10.1056/NEJMoa0907847
-
6. Ohno H, Koga H, Kohno S, Tashiro K, Hara K. Relationship between rifampin MICs for and rpoB mutations of Mycobacterium tuberculosis strains isolated in Japan. Antimicrob Agents Chemother. 1996;40(5):1053–1056.
-
7. Heep M, Rieger U, Beck D, Lehn N. Mutations in the beginning of the rpoB gene can induce resistance to rifamycins
in both Helicobacter pylori and Mycobacterium tuberculosis. Antimicrob Agents Chemother. 2000;44(4):1075–1077.
-
8. Taniguchi H, Aramaki H, Nikaido Y, Mizuguchi Y, Nakamura M, Koga T, et al. Rifampicin resistance and mutation of the rpoB gene in Mycobacterium tuberculosis. FEMS Microbiol Lett. 1996;144(1):103–108.
-
9. Lee PH, Chan PC, Peng YT, Chu PW, Wu MH, Jou R, et al. Impact of universal drug susceptibility testing and effective management of multidrug-resistant tuberculosis in Taiwan. PLoS One. 2019;14(4):e0214792.
-
10. Yu MC, Chiang CY, Lee JJ, Chien ST, Lin CJ, Lee SW, et al. Treatment outcomes of multidrug-resistant tuberculosis in Taiwan: tackling loss to follow-up. Clin Infect Dis. 2018;67(2):202–210. doi:10.1093/cid/ciy066
-
11. Chan PC, Huang SH, Yu MC, Lee SW, Huang YW, Chien ST, et al. Effectiveness of a government-organized and hospital-initiated treatment for multidrug-resistant tuberculosis patients—a retrospective cohort study. PLoS One. 2013;8(2):e57719. doi:10.1371/journal.pone.0057719
-
12. Georghiou SB, Gomathi NS, Rajendran P, Nagalakshmi V, Prabakaran L, Prem Kumar MM, et al. Accuracy of the Truenat MTB-RIF Dx assay for detection of rifampicin resistance-associated mutations. Tuberculosis (Edinb). 2021;127:102064.
-
13. Singh UB, Singh M, Sharma S, Mahajan N, Bala K, Srivastav A, et al. Expedited diagnosis of pediatric tuberculosis using Truenat MTB-Rif Dx and GeneXpert MTB/RIF. Sci Rep. 2023;13:6976.
-
14. Meaza A, Tesfaye E, Mohamed Z, Zerihun B, Seid G, Eshetu K, et al. Diagnostic accuracy of Truenat Tuberculosis and Rifampicin-Resistance assays in Addis Ababa, Ethiopia. PLoS One. 2021;16(5):e0261084.
-
15. Omer ZB, Mekonnen Y, Worku A, Zewde A, Medhin G, Mohammed T, et al. Evaluation of the GenoType MTBDRplus assay for detection of rifampicin-and isoniazid-resistant Mycobacterium tuberculosis isolates in central Ethiopia. Int J Mycobacteriol. 2016;5(4):475–481.
-
16. Siddiqui S, Brooks MB, Malik AA, Fuad J, Nazish A, Bano S, et al. Evaluation of GenoType MTBDR plus for the detection of drug-resistant Mycobacterium tuberculosis on isolates from Karachi, Pakistan. PLoS One. 2019;14(8):e0221485.
-
17. Meaza A, Kebede A, Yaregal Z, Dagne Z, Moga S, Yenew B, et al. Evaluation of genotype MTBDRplus VER 2.0 line probe assay for the detection of MDR-TB in smear positive and negative sputum samples. BMC Infect Dis. 2017;17(1):280.
-
18. Abanda NN, Djieugoué JY, Lim E, Pefura-Yone EW, Mbacham WF, Vernet G, et al. Diagnostic accuracy and usefulness of the Genotype MTBDRplus assay in diagnosing multidrug-resistant tuberculosis in Cameroon: A cross-sectional study. BMC Infect Dis. 2017;17(1):379 doi: 10.1186/s12879-017-2489-3.
-
19. Jian J, Yang X, Yang J, Chen L. Evaluation of the GenoType MTBDRplus and MTBDRsl for the detection of drug-resistant Mycobacterium tuberculosis on isolates from Beijing, China. Infect Drug Resist. 2018;11:1627–1634.
-
20. Singh BK, Sharma R, Kodan P, Soneja M, Jorwal P, Nischal N, et al. Diagnostic evaluation of non-interpretable results associated with rpoB gene in Genotype MTBDRplus Ver 2.0. Tuberc Respir Dis (Seoul). 2020;83(4):289–294.
-
21. Maurya AK, Singh AK, Kant S, Umrao J, Kumar M, Kushwaha RAS, et al. Use of GenoType® MTBDRplus assay to assess drug resistance and mutation patterns of multidrug-resistant tuberculosis isolates in northern India. Indian J Med Microbiol. 2013;31(3):230–236.
Moleküler Yöntemlerle İlaç Dirençli Mycobacterium tuberculosis Suşlarının Araştırılması
Yıl 2025,
Cilt: 51 Sayı: 2, 291 - 294, 28.08.2025
Ayşe Melda Payaslıoğlu
,
Sezcan Sağlam
,
Seçil Ak Aksoy
,
Cüneyt Özakın
Öz
Bu çalışmanın amacı, fenotipik olarak rifampisin ve/veya etambutol dirençli Mycobacterium tuberculosis izolatlarında, rpoB ve embB genlerindeki mutasyonları moleküler yöntemlerle araştırmaktır. 2005–2014 yılları arasında Türkiye’den toplanan 110 izolatın ilaç duyarlılıkları modifiye agar proporsiyon yöntemiyle test edilmiştir. Seçilen 28 suşta moleküler analiz yapılmış, 7’si rifampisin dirençli bulunmuştur. rpoB geninde, Rifampisin Direnç Belirleyici Bölge (RRDR) olan 507–533. kodonlar arasında mutasyonlar tespit edilmiştir. En yaygın mutasyon, 10 suşta görülen Ser531Leu (TCG→TTG) mutasyonu, bir suşta ise His526Asn (CAC→AAC) mutasyonudur. Bu mutasyonlar, rifampisin direncinin moleküler göstergeleri olarak önemli bulunmuştur. Moleküler yöntemlerle bu mutasyonların hızlı tespiti, tüberküloz tedavisinin erken planlanması için önemlidir.
Kaynakça
-
1. Gagneux S. Ecology and evolution of Mycobacterium tuberculosis. Nat Rev Microbiol. 2018;16(4):202–213. doi:10.1038/nrmicro.2018.8
-
2. Pai M, Behr MA, Dowdy D, Dheda K, Divangahi M, Boehme CC, et al. Tuberculosis. Nat Rev Dis Primers. 2016;2:16076. doi:10.1038/nrdp.2016.76
-
3. World Health Organization. Global Tuberculosis Report 2023. 2023. [Accessed: June 23, 2024]
-
4. Dong B, He Z, Li Y, Xu X, Wang C, Zeng J. Improved conventional and new approaches in the diagnosis of tuberculosis. Front Microbiol. 2022;13:924410.
-
5. Boehme CC, Nabeta P, Hillemann D, Nicol MP, Shenai S, Krapp F, et al. Rapid molecular detection of tuberculosis and rifampin resistance. N Engl J Med. 2010;363(11):1005–1015. doi:10.1056/NEJMoa0907847
-
6. Ohno H, Koga H, Kohno S, Tashiro K, Hara K. Relationship between rifampin MICs for and rpoB mutations of Mycobacterium tuberculosis strains isolated in Japan. Antimicrob Agents Chemother. 1996;40(5):1053–1056.
-
7. Heep M, Rieger U, Beck D, Lehn N. Mutations in the beginning of the rpoB gene can induce resistance to rifamycins
in both Helicobacter pylori and Mycobacterium tuberculosis. Antimicrob Agents Chemother. 2000;44(4):1075–1077.
-
8. Taniguchi H, Aramaki H, Nikaido Y, Mizuguchi Y, Nakamura M, Koga T, et al. Rifampicin resistance and mutation of the rpoB gene in Mycobacterium tuberculosis. FEMS Microbiol Lett. 1996;144(1):103–108.
-
9. Lee PH, Chan PC, Peng YT, Chu PW, Wu MH, Jou R, et al. Impact of universal drug susceptibility testing and effective management of multidrug-resistant tuberculosis in Taiwan. PLoS One. 2019;14(4):e0214792.
-
10. Yu MC, Chiang CY, Lee JJ, Chien ST, Lin CJ, Lee SW, et al. Treatment outcomes of multidrug-resistant tuberculosis in Taiwan: tackling loss to follow-up. Clin Infect Dis. 2018;67(2):202–210. doi:10.1093/cid/ciy066
-
11. Chan PC, Huang SH, Yu MC, Lee SW, Huang YW, Chien ST, et al. Effectiveness of a government-organized and hospital-initiated treatment for multidrug-resistant tuberculosis patients—a retrospective cohort study. PLoS One. 2013;8(2):e57719. doi:10.1371/journal.pone.0057719
-
12. Georghiou SB, Gomathi NS, Rajendran P, Nagalakshmi V, Prabakaran L, Prem Kumar MM, et al. Accuracy of the Truenat MTB-RIF Dx assay for detection of rifampicin resistance-associated mutations. Tuberculosis (Edinb). 2021;127:102064.
-
13. Singh UB, Singh M, Sharma S, Mahajan N, Bala K, Srivastav A, et al. Expedited diagnosis of pediatric tuberculosis using Truenat MTB-Rif Dx and GeneXpert MTB/RIF. Sci Rep. 2023;13:6976.
-
14. Meaza A, Tesfaye E, Mohamed Z, Zerihun B, Seid G, Eshetu K, et al. Diagnostic accuracy of Truenat Tuberculosis and Rifampicin-Resistance assays in Addis Ababa, Ethiopia. PLoS One. 2021;16(5):e0261084.
-
15. Omer ZB, Mekonnen Y, Worku A, Zewde A, Medhin G, Mohammed T, et al. Evaluation of the GenoType MTBDRplus assay for detection of rifampicin-and isoniazid-resistant Mycobacterium tuberculosis isolates in central Ethiopia. Int J Mycobacteriol. 2016;5(4):475–481.
-
16. Siddiqui S, Brooks MB, Malik AA, Fuad J, Nazish A, Bano S, et al. Evaluation of GenoType MTBDR plus for the detection of drug-resistant Mycobacterium tuberculosis on isolates from Karachi, Pakistan. PLoS One. 2019;14(8):e0221485.
-
17. Meaza A, Kebede A, Yaregal Z, Dagne Z, Moga S, Yenew B, et al. Evaluation of genotype MTBDRplus VER 2.0 line probe assay for the detection of MDR-TB in smear positive and negative sputum samples. BMC Infect Dis. 2017;17(1):280.
-
18. Abanda NN, Djieugoué JY, Lim E, Pefura-Yone EW, Mbacham WF, Vernet G, et al. Diagnostic accuracy and usefulness of the Genotype MTBDRplus assay in diagnosing multidrug-resistant tuberculosis in Cameroon: A cross-sectional study. BMC Infect Dis. 2017;17(1):379 doi: 10.1186/s12879-017-2489-3.
-
19. Jian J, Yang X, Yang J, Chen L. Evaluation of the GenoType MTBDRplus and MTBDRsl for the detection of drug-resistant Mycobacterium tuberculosis on isolates from Beijing, China. Infect Drug Resist. 2018;11:1627–1634.
-
20. Singh BK, Sharma R, Kodan P, Soneja M, Jorwal P, Nischal N, et al. Diagnostic evaluation of non-interpretable results associated with rpoB gene in Genotype MTBDRplus Ver 2.0. Tuberc Respir Dis (Seoul). 2020;83(4):289–294.
-
21. Maurya AK, Singh AK, Kant S, Umrao J, Kumar M, Kushwaha RAS, et al. Use of GenoType® MTBDRplus assay to assess drug resistance and mutation patterns of multidrug-resistant tuberculosis isolates in northern India. Indian J Med Microbiol. 2013;31(3):230–236.