Molecular Identification of non-tuberculous mycobacteria
Keywords:
Mycobacterium infections, atypical, polymerase chain reaction, molecular diagnostictechniques, opportunistic infections, public health
Abstract
Introduction. Nontuberculous mycobacteria can be saprophytic, pathogenic or opportunistic. The most common diseases produced by these microorganisms are the post-surgical infections due to anesthetic procedures, infections associated with catheters, disseminated cutaneous diseases and pulmonary and central nervous system diseases that especially affect HIV patients. Identification of the nontuberculous mycobacteria can take several weeks and even then, differentiation of complex members is not possible.Objective. The PCR-restriction analysis (PRA) technique was evaluated as a method for genotypic identification of nontuberculous mycobacteria isolated of clinical samples located in the culture collection of the Instituto Nacional de Salud (National Institute of Health), Bogotá, Colombia.
Materials and methods. Seventy clinical isolates of nontuberculous mycobacteria stored in 50% glycerol at -70°C were identified by phenotypic techniques. The genotypic identification was made using the PCR-restriction analysis (PRA) using the restriction enzymes BstEII and HseIII, the restriction products were visualized on gels of agarose to 3%, and the concordance between the methodologies was evaluated.
Results. A matching of 100% was obtained in the identification of Mycobacterium terrae, M. szulgai, M. avium, M. chelonae and M. scrofulaceum, the matching between M. fortuitum species, M. abscessus, M. gordonae and M. intracellulare varied from 44 to 89%; there was no concurrence in the identification of species M. flavescens and M. malmoense.
Conclusions. PRA provided a fast, inexpensive and accurate alternative for the identification of nontuberculous mycobacteria that permited the differentiation among species of a complex and determining the subtype of each species sample.
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References
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27. Garzón MC, Naranjo ON, Llerena C, Orjuela DL. Identificación de las micobacterias. Manual de procedimientos. Bogotá: Instituto Nacional de Salud; 2002.
28. Springer B, Stockman L, Teschner K, Roberts GD, Bottger EC. Two laboratory collaborative study on identification of mycobacteria: molecular versus phenotypic methods. J Clin Microbiol. 1996;34:296-303.
29. Byrd TF, Lyons CR. Preliminary characterization of a Mycobacterium abscessus mutant in human and murine models of infection. Infect Immun. 1999;67:4700-7.
30. Maekura R, Okuda Y, Hirotani A, Kitada S, Hiraga T, Yoshimura K, et al. Clinical and prognostic importance of serotyping Mycobacterium avium-Mycobacterium intracellulare complex isolates in human immunodeficiency virus-negative patients. J Clin Microbiol. 2005;43:3150-8.
31. García LM, Garzón M, Orjuela D, Mejía G, Llerena C, Angee D. Mycobacterium tuberculosis y micobacterias no tuberculosas en procedimientos de mesoterapia. Inf Quinc Epidemiol Nac. 2007;12:40-4.
2. Primm TP, Lucero CA, Falkinham JO 3rd. Health impacts of environmental mycobacteria. Clin Microbiol Rev. 2004;17:98-106.
3. Tortoli E. Impact of genotypic studies on mycobacterial taxonomy: the new mycobacteria of the 1990s. Clin Microbiol Rev. 2003;16:319-54.
4. Nuñez-Guzman J, Starke J, Correa A, Graviss E, Pan X, Popek E, et al. A report of cutaneous tuberculosis in siblings. Pediatr Dermatol. 2003;20:404-7.
5. Duttaroy B, Agrawal C, Pendse A. Spinal tuberculosis due to dissemination of atypical mycobacteria. Indian J Med Sci. 2004;58:203-5.
6. Holmes GF, Harrington SM, Romagnoli MJ, Merz WG. Recurrent, disseminated Mycobacterium marinum infection caused by the same genotypically defined strain in an immunocompromised patient. J Clin
Microbiol. 1999;37:3059-61.
7. Murcia-Aranguren MI, Gómez-Marín JE, Alvarado FS, Bustillo JG, de Mendivelson E, Gomez B, et al. Frequency of tuberculous and non-tuberculous mycobacteria in HIV infected patients from Bogotá,
Colombia. BMC Infect Dis. 2001;1:21.
8. Camargo D, Saad C, Ruiz F, Ramírez ME, Lineros M, Rodríguez G, et al. Iatrogenic outbreak of M. chelonae skin abscesses. Epidemiol Infect. 1996;177:113-9.
9. Ortegón M, Rodríguez G, Camargo D, Orozco LC. Mycobacterium chelonae y Mycobacterium abscessus: patógenos emergentes. Biomédica. 1996;16:217-38.
10. Garzón M, Orjuela D, Naranjo O, Llerena C. Micobacterias no tuberculosas en Colombia 1995-2003. Inf Quinc Epidemiol Nac. 2005;10:161-76.
11. León CI, Jiménez Y, López OJ, Pavas J, Bustillo JG, Guerrero MI. Evolución de la asociación micobacteriana HIV-SIDA en Bogotá 1995-2003. Infectio. 2004;8:99.
12. Crespo M, Corral R, Alzate A. Micobacterias no tuberculosas en personas VIH positivas y en personas sin factores de riesgo a la infección. Colombia Med. 1994;25:86-91.
13. León CI. Presencia de las micobacterias no tuberculosas en Colombia. Médicas UIS. 1998;12:181-7.
14. Wang SX, Tay L, Sing LH. Rapid identification of pathogenic rapidly growing mycobacteria by PCR-restriction endonuclease analysis. Ann Acad Med Singapore. 2005;34:137-40.
15. Vélez E. Actividad in vitro de diferentes antimicobacterianos frente a cepas de micobacterias no tuberculosas aisladas en Colombia (tesis). Bogotá:Universidad Nacional de Colombia; 1999.
16. Telenti A, Marchesi F, Balz M, Bally F, Bötter EC, Bodmer T. Rapid identification of mycobacteria to the species level by polymerase chain reaction and restriction enzyme analysis. J Clin Microbiol. 1993;31:175-8.
17. Da Silva C, Misuka S, Cássia D, Cardoso S. Hsp65 PCR-restriction enzyme analysis (PRA) for identification of mycobacteria in the clinical laboratory. Rev Inst Med Trop Sao Paulo. 2001;43:25-8.
18. Devallois A, Goh KS, Rastogi N. Rapid identification of mycobacteria to species level by PCR-restriction fragment length polymorphism analysis of the hsp65 gene and proposition of an algorithm to differentiate 34 mycobacteria species. J Clin Microbiol. 1997;35:2969-73.
19. Brunello F, Ligozzi M, Cristelli E, Bonora S, Tortoli E, Fontana R. Identification of 54 mycobacterial species by PCR-restriction fragment length polymorphism analysis of the hsp65 gene. J Clin Microbiol. 2001;39:2799-806.
20. Rastogi N, Goh KS, Berchel M. Species-specific identification of Mycobacterium leprae by PCR-restriction fragment length polymorphism analysis of the hsp65 gene. J Clin Microbiol. 1999;37:2016-9.
21. Leao SC, Bernardelli A, Cataldi A, Zumarraga M, Robledo J, Realpe T, et al. Multicenter evaluation of mycobacteria identification by PCR restriction enzyme analysis in laboratories from Latin America and the Caribbean. J Microbiol Methods. 2005;61:193-9.
22. Murcia MI, Tortoli E, Menéndez MC, Palenque E, García MJ. Mycobacterium colombiense sp. nov., a novel member of the Mycobacterium avium complex and description of MAC-X as a new ITS genetic variant. Int J Syst Evol Microbiol. 2006;56:2049-54.
23. Tortoli E, Rindi L, Garcia MJ, Chiaradonna P, Dei R, Garzelli C, et al. Proposal to elevate the genetic variant MAC-A, included in the Mycobacterium avium complex, to species rank as Mycobacterium chimaera sp nov. Int J Syst Evol Microbiol. 2004;54:1277-85
24. Leao S, Martn A, Mejía G, Palomino J, Robledo J, Telles Da Silva M, et al. Practical handbook for phenotypic and genotypic identification of Micobacteria. First edition. Brussels: European Commision, International Cooperation for Developing Countries; 2004.
25. Garzón MC, Naranjo ON, Sierra CR, Llerena C, Orjuela DL. Bacteriología del Mycobacterium tuberculosis y de micobacterias no tuberculosas. Manual de Procedimientos. 1ª edición. Bogotá: Instituto Nacional de Salud; 2001.
26. Van Soolingen D, Hermans PW, de Haas PE, Soll DR, van Embden JD. The occurrence and stability of insertion sequences in Mycobacterium tuberculosis complex strains; evaluation of IS-dependent DNA polymorphism as a tool in the epidemiology of tuberculosis. J Clin Microbiol. 1991;29:2578-86.
27. Garzón MC, Naranjo ON, Llerena C, Orjuela DL. Identificación de las micobacterias. Manual de procedimientos. Bogotá: Instituto Nacional de Salud; 2002.
28. Springer B, Stockman L, Teschner K, Roberts GD, Bottger EC. Two laboratory collaborative study on identification of mycobacteria: molecular versus phenotypic methods. J Clin Microbiol. 1996;34:296-303.
29. Byrd TF, Lyons CR. Preliminary characterization of a Mycobacterium abscessus mutant in human and murine models of infection. Infect Immun. 1999;67:4700-7.
30. Maekura R, Okuda Y, Hirotani A, Kitada S, Hiraga T, Yoshimura K, et al. Clinical and prognostic importance of serotyping Mycobacterium avium-Mycobacterium intracellulare complex isolates in human immunodeficiency virus-negative patients. J Clin Microbiol. 2005;43:3150-8.
31. García LM, Garzón M, Orjuela D, Mejía G, Llerena C, Angee D. Mycobacterium tuberculosis y micobacterias no tuberculosas en procedimientos de mesoterapia. Inf Quinc Epidemiol Nac. 2007;12:40-4.
How to Cite
1.
Castro CM, Puerto G, García LM, Orjuela DL, Llerena C, Garzón MC, et al. Molecular Identification of non-tuberculous mycobacteria. biomedica [Internet]. 2007 Sep. 1 [cited 2024 May 18];27(3):439-46. Available from: https://revistabiomedica.org/index.php/biomedica/article/view/206
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