Genotyping of Mycobacterium leprae strains in south central coast and central highlands of Vietnam
Abstract
Background and Objectives: Leprosy remains an important health problem worldwide. It is one of the oldest recorded diseases of humankind. In this study, we expanded the analysis of the geographic distribution of Mycobacterium leprae by investigating SNPs and rpoT genotypes in South Central Coast and Central Highlands clinical isolates, providing insights into the distribution and transmission of leprosy in Vietnam and in this geographic region.
Materials and Methods: 27 clinical isolates from the patients, determined the genotypes of M. leprae by SNP and rpoT polymorphism. SNP genotyping was performed by PCR amplification and sequencing, rpoT genotyping by PCR amplification and electrophoresis.
Results: All of 27 DNA samples (100%) were positive with RLEP TaqMan PCR (Ct value range is 18-32 on 3 replicates). SNP type 1 was identified in 15 isolates (56%), while SNP type 3 was detected in 12 samples (44%). SNP type 2 and type 4, were not detected. The 6-base repeat region of the rpoT gene was amplified by PCR and analyzed by 4% MetaPhor™ agarose gel electrophoresis. All isolates yielded amplification products of 91-bp, but not 97-bp.
Conclusion: This study showed that 56% of isolates belonged to type 1, 44% to type 3. In addition, all samples have the 3-copy hexamer genotype in the rpoT gene.
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5. Wang L, Sun PW, Yu MW, Gu H, Wang H-S, Chen XS. Epidemiological characteristics of leprosy in China, 2018. Int J Dermatol Venereol 2020; 3: 27-30.
6. Wang L, Sun PW, Yu MW, Gu H, Wang HS, Chen XS. Leprosy update in China, 2019. Int J Dermatol Venereol 2022; 5: 15-19.
7. Chae GT. Modern history of Hansen’s disease in Korea. Infect Chemother 2020; 52: 647-653.
8. Rao Pn, Suneetha S. Current situation of leprosy in India and its future implications. Indian Dermatol Online J 2018; 9: 83-89.
9. Chen K-H, Lin C-Y, Su S-B, Chen K-T. Leprosy: A review of epidemiology, clinical diagnosis, and management. J Trop Med 2022; 2022: 8652062.
10. Truman RW, Singh P, Sharma R, Busso P, Rougemont J, Paniz-Mondolfi A, et al. Probable Zoonotic Leprosy in the Southern United States. N Engl J Med 2011; 364: 1626-1633.
11. Monot M, Honoré N, Balière C, Ji B, Sow S, Brennan PJ, et al. Are variable-number tandem repeats appropriate for Genotyping Mycobacterium leprae ? J Clin Microbiol 2008; 46: 2291-2297.
12. Salipante SJ, Hall BG. Towards the molecular epidemiology of Mycobacterium leprae: Strategies, successes, and shortcomings. Infect Genet Evol 2011; 11: 1505-1513.
13. Das M, Chaitanya VS, Kanmani K, Rajan L, Ebenezer M. Genomic diversity in Mycobacterium leprae isolates from leprosy cases in South India. Infect Genet Evol 2016; 45: 285-289.
14. Monot M, Honoré N, Garnier T, Araoz R, Coppée J-Y, Lacroix C, et al. On the origin of leprosy. Science 2005; 308: 1040-1042.
15. Shin YC, Lee H, Lee H, Walsh GP, Kim JD, Cho SN. Variable numbers of TTC repeats in Mycobacterium leprae DNA from leprosy patients and use in strain differentiation. J Clin Microbiol 2000; 38: 4535-4538.
16. Matsuoka M, Zhang L, Morris MF, Legua P, Wiens C. Polymorphism in the rpoT gene in Mycobacterium leprae isolates obtained from Latin American countries and its possible correlation with the spread of leprosy. FEMS Microbiol Lett 2005; 243: 311-315.
17. Matsuoka M, Lopez Roa RI, Budiawan T, Kyaw K, Chae GT. Genotypic analysis of Mycobacterium leprae isolates from Japan and other Asian countries reveals a global transmission pattern of leprosy. FEMS Microbiol Lett 2006; 261: 150-154.
18. Groathouse NA, Rivoire B, Kim H, Lee H, Cho S-N, Brennan PJ, et al. Multiple polymorphic loci for molecular typing of strains of Mycobacterium leprae. J Clin Microbiol 2004; 42: 1666-1672.
19. Truman R, Fontes AB, De Miranda AB, Suffys P, Gillis T. Genotypic variation and stability of four variable-number tandem repeats and their suitability for discriminating strains of Mycobacterium leprae. J Clin Microbiol 2004; 42: 2558-2565.
20. Monot M, Honoré N, Garnier T, Zidane N, Sherafi D, Paniz-Mondolfi A, et al. Comparative genomic and phylogeographic analysis of Mycobacterium leprae. Nat Genet 2009; 41: 1282-1289.
21. Kai M, Nguyen Phuc NH, Nguyen HA, Pham TH, Nguyen KH, Miyamoto Y, et al. Analysis of drug-resistant strains of Mycobacterium leprae in an endemic area of Vietnam. Clin Infect Dis 2011; 52(5): e127-132.
22. Truman RW, Andrews PK, Robbins NY, Adams LB, Krahenbuhl JL, Gillis TP. Enumeration of Mycobacterium leprae using real-time PCR. PLoS Negl Trop Dis 2008; 2(11): e328.
23. Phetsuksiri B, Srisungngam S, Rudeeaneksin J, Bunchoo S, Lukebua A, Wongtrungkapun R, et al. SNP genotypes of Mycobacterium leprae isolates in Thailand and their combination with rpoT and TTC genotyping for analysis of leprosy distribution and transmission. Jpn J Infect Dis 2012; 65: 52–56.
24. Weng X, Wang Z, Liu J, Kimura M, Black WC 4th, Brennan PJ, et al. Identification and distribution of Mycobacterium leprae genotypes in a region of high leprosy prevalence in China: A 3-year molecular epidemiological study. J Clin Microbiol 2007; 45: 1728-1734.
25. Yuan Y, Wen Y, You Y, Xing Y, Li H, Weng X, et al. Characterization of Mycobacterium leprae genotypes in China-identification of a new polymorphism C251T in the 16S rRNA gene. PLoS One 2015; 10(7): e0133268.
26. Sakamuri RM, Kimura M, Li W, Kim HC, Lee H, Kiran MD, et al. Population-based molecular epidemiology of leprosy in Cebu, Philippines. J Clin Microbiol 2009; 47: 2844-2854.
27. Chae GT, Lee SB, Kang TJ, Shin HK, Kim JP, Ko YH, et al. Typing of clinical isolates of Mycobacterium leprae and their distribution in Korea. Lepr Rev 2002; 73: 41-46.
28. Lavania M, Lal R, Joseph G, Darlong J, Abraham S, Nanda NK, et al. Genotypic analysis of Mycobacterium leprae strains from different regions of India on the basis of rpoT. Indian J Lepr 2009; 81: 119-124.
29. Lavania M, Katoch K, Singh H, Das R, Gupta AK, Sharma R, et al. Predominance of three copies of tandem repeats in rpoT gene of Mycobacterium leprae from Northern India. Infect Genet Evol 2007; 7: 627-631.
2. World Health Organization. Global leprosy (Hansen disease) update, 2021: moving towards interruption of transmission. 2021; Retrieved from https://www.who.int/publications/i/item/who-wer9736-429-450
3. World Health Organization. Leprosy (Hansen’s disease). 2021; Retrieved from: https://www.who.int/data/gho/data/themes/topics/leprosy-hansens-disease
4. World Health Organization. Regional Office for the Western Pacific. (2011). Epidemiological review of leprosy in the Western Pacific Region 2008-2010: sustaining leprosy services and further reducing the leprosy burden. WHO Regional Office for the Western Pacific. Retrieved from https://apps.who.int/iris/handle/10665/207500
5. Wang L, Sun PW, Yu MW, Gu H, Wang H-S, Chen XS. Epidemiological characteristics of leprosy in China, 2018. Int J Dermatol Venereol 2020; 3: 27-30.
6. Wang L, Sun PW, Yu MW, Gu H, Wang HS, Chen XS. Leprosy update in China, 2019. Int J Dermatol Venereol 2022; 5: 15-19.
7. Chae GT. Modern history of Hansen’s disease in Korea. Infect Chemother 2020; 52: 647-653.
8. Rao Pn, Suneetha S. Current situation of leprosy in India and its future implications. Indian Dermatol Online J 2018; 9: 83-89.
9. Chen K-H, Lin C-Y, Su S-B, Chen K-T. Leprosy: A review of epidemiology, clinical diagnosis, and management. J Trop Med 2022; 2022: 8652062.
10. Truman RW, Singh P, Sharma R, Busso P, Rougemont J, Paniz-Mondolfi A, et al. Probable Zoonotic Leprosy in the Southern United States. N Engl J Med 2011; 364: 1626-1633.
11. Monot M, Honoré N, Balière C, Ji B, Sow S, Brennan PJ, et al. Are variable-number tandem repeats appropriate for Genotyping Mycobacterium leprae ? J Clin Microbiol 2008; 46: 2291-2297.
12. Salipante SJ, Hall BG. Towards the molecular epidemiology of Mycobacterium leprae: Strategies, successes, and shortcomings. Infect Genet Evol 2011; 11: 1505-1513.
13. Das M, Chaitanya VS, Kanmani K, Rajan L, Ebenezer M. Genomic diversity in Mycobacterium leprae isolates from leprosy cases in South India. Infect Genet Evol 2016; 45: 285-289.
14. Monot M, Honoré N, Garnier T, Araoz R, Coppée J-Y, Lacroix C, et al. On the origin of leprosy. Science 2005; 308: 1040-1042.
15. Shin YC, Lee H, Lee H, Walsh GP, Kim JD, Cho SN. Variable numbers of TTC repeats in Mycobacterium leprae DNA from leprosy patients and use in strain differentiation. J Clin Microbiol 2000; 38: 4535-4538.
16. Matsuoka M, Zhang L, Morris MF, Legua P, Wiens C. Polymorphism in the rpoT gene in Mycobacterium leprae isolates obtained from Latin American countries and its possible correlation with the spread of leprosy. FEMS Microbiol Lett 2005; 243: 311-315.
17. Matsuoka M, Lopez Roa RI, Budiawan T, Kyaw K, Chae GT. Genotypic analysis of Mycobacterium leprae isolates from Japan and other Asian countries reveals a global transmission pattern of leprosy. FEMS Microbiol Lett 2006; 261: 150-154.
18. Groathouse NA, Rivoire B, Kim H, Lee H, Cho S-N, Brennan PJ, et al. Multiple polymorphic loci for molecular typing of strains of Mycobacterium leprae. J Clin Microbiol 2004; 42: 1666-1672.
19. Truman R, Fontes AB, De Miranda AB, Suffys P, Gillis T. Genotypic variation and stability of four variable-number tandem repeats and their suitability for discriminating strains of Mycobacterium leprae. J Clin Microbiol 2004; 42: 2558-2565.
20. Monot M, Honoré N, Garnier T, Zidane N, Sherafi D, Paniz-Mondolfi A, et al. Comparative genomic and phylogeographic analysis of Mycobacterium leprae. Nat Genet 2009; 41: 1282-1289.
21. Kai M, Nguyen Phuc NH, Nguyen HA, Pham TH, Nguyen KH, Miyamoto Y, et al. Analysis of drug-resistant strains of Mycobacterium leprae in an endemic area of Vietnam. Clin Infect Dis 2011; 52(5): e127-132.
22. Truman RW, Andrews PK, Robbins NY, Adams LB, Krahenbuhl JL, Gillis TP. Enumeration of Mycobacterium leprae using real-time PCR. PLoS Negl Trop Dis 2008; 2(11): e328.
23. Phetsuksiri B, Srisungngam S, Rudeeaneksin J, Bunchoo S, Lukebua A, Wongtrungkapun R, et al. SNP genotypes of Mycobacterium leprae isolates in Thailand and their combination with rpoT and TTC genotyping for analysis of leprosy distribution and transmission. Jpn J Infect Dis 2012; 65: 52–56.
24. Weng X, Wang Z, Liu J, Kimura M, Black WC 4th, Brennan PJ, et al. Identification and distribution of Mycobacterium leprae genotypes in a region of high leprosy prevalence in China: A 3-year molecular epidemiological study. J Clin Microbiol 2007; 45: 1728-1734.
25. Yuan Y, Wen Y, You Y, Xing Y, Li H, Weng X, et al. Characterization of Mycobacterium leprae genotypes in China-identification of a new polymorphism C251T in the 16S rRNA gene. PLoS One 2015; 10(7): e0133268.
26. Sakamuri RM, Kimura M, Li W, Kim HC, Lee H, Kiran MD, et al. Population-based molecular epidemiology of leprosy in Cebu, Philippines. J Clin Microbiol 2009; 47: 2844-2854.
27. Chae GT, Lee SB, Kang TJ, Shin HK, Kim JP, Ko YH, et al. Typing of clinical isolates of Mycobacterium leprae and their distribution in Korea. Lepr Rev 2002; 73: 41-46.
28. Lavania M, Lal R, Joseph G, Darlong J, Abraham S, Nanda NK, et al. Genotypic analysis of Mycobacterium leprae strains from different regions of India on the basis of rpoT. Indian J Lepr 2009; 81: 119-124.
29. Lavania M, Katoch K, Singh H, Das R, Gupta AK, Sharma R, et al. Predominance of three copies of tandem repeats in rpoT gene of Mycobacterium leprae from Northern India. Infect Genet Evol 2007; 7: 627-631.
| Files | ||
| Issue | Vol 15 No 2 (2023) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v15i2.12470 | |
| Keywords | ||
| Leprosy; rpoT protein; Mycobacterium leprae; Pseudogenes | ||
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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
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(2):201-207.
