Utilization of molecular and conventional methods for the identification of nontuberculous mycobacteria isolated from different water sources
Background: The environment is the nontuberculous mycobacteria (NTM) reservoir, opportunistic pathogens of great diversity and ubiquity, which is observed in the constant description of new species capable of causing infection. Since its introduction, molecular methods are essential for species iden...
| Autores principales: | , , , |
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| Formato: | info:ar-repo/semantics/artículo |
| Lenguaje: | Inglés |
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Wolters Kluwer - Medknow
2018
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| Materias: | |
| Acceso en línea: | http://www.ijmyco.org/article.asp?issn=2212-5531;year=2018;volume=7;issue=1;spage=53;epage=60;aulast=Tortone http://hdl.handle.net/20.500.12123/3625 |
| _version_ | 1855035168771801088 |
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| author | Tortone, Claudia Andrea Zumarraga, Martin Jose Gioffre, Andrea Oriani, Delia Susana |
| author_browse | Gioffre, Andrea Oriani, Delia Susana Tortone, Claudia Andrea Zumarraga, Martin Jose |
| author_facet | Tortone, Claudia Andrea Zumarraga, Martin Jose Gioffre, Andrea Oriani, Delia Susana |
| author_sort | Tortone, Claudia Andrea |
| collection | INTA Digital |
| description | Background: The environment is the nontuberculous mycobacteria (NTM) reservoir, opportunistic pathogens of great diversity and ubiquity, which is observed in the constant description of new species capable of causing infection. Since its introduction, molecular methods are essential for species identification. Most comparative studies between molecular and conventional methods, have used isolated strains from clinical samples. Methods: The aim of this study was to evaluate the usefulness of molecular methods, especially the hsp65-PRA (PCR-Restriction Enzyme Analysis), and biochemical tests in the identification of NTM recovered from water of different origins, using the sequencing of 16S rRNA and hsp 65 genes as assessment methods of the previous ones. Species identification was performed for all 56 NTM isolates what were recovered from 32 (42.1%) positive water samples, using conventional phenotypic methods, hsp65-PRA, partial sequencing of 16S rRNA and sequencing of hsp 65 genes. Results: Phenotypic evaluation and hsp65-PRA were concordant with 23 (41.1%) isolates. Also, the PRA was concordant with 16 (28.6%) and 27 (48.2%) isolates, with the partial sequencing of 16S rRNA and sequencing of hsp 65 genes, respectively. It is considered that the 19.6% (n = 11) could not be identified. Conclusion: Identification of NTM environmental isolates to the species level, especially when they are pigmented and fast-growing, both the analysis of the restriction patterns obtained by PRA and the sequencing of the 16S rRNA and hsp 65 genes are insufficient by themselves. Although they are demanding and time-consuming, biochemical tests are very useful to support data obtained by molecular methods. |
| format | info:ar-repo/semantics/artículo |
| id | INTA3625 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2018 |
| publishDateRange | 2018 |
| publishDateSort | 2018 |
| publisher | Wolters Kluwer - Medknow |
| publisherStr | Wolters Kluwer - Medknow |
| record_format | dspace |
| spelling | INTA36252018-10-18T12:55:29Z Utilization of molecular and conventional methods for the identification of nontuberculous mycobacteria isolated from different water sources Tortone, Claudia Andrea Zumarraga, Martin Jose Gioffre, Andrea Oriani, Delia Susana Enzymatic Analysis Waterborne Diseases Análisis Enzimático Enfermedades Transmitidas por Agua Nontuberculous Mycobacteria Phenotypic Methods Water Sources Micobacterias no Tuberculosas Métodos Fenotípicos Fuentes de Agua Background: The environment is the nontuberculous mycobacteria (NTM) reservoir, opportunistic pathogens of great diversity and ubiquity, which is observed in the constant description of new species capable of causing infection. Since its introduction, molecular methods are essential for species identification. Most comparative studies between molecular and conventional methods, have used isolated strains from clinical samples. Methods: The aim of this study was to evaluate the usefulness of molecular methods, especially the hsp65-PRA (PCR-Restriction Enzyme Analysis), and biochemical tests in the identification of NTM recovered from water of different origins, using the sequencing of 16S rRNA and hsp 65 genes as assessment methods of the previous ones. Species identification was performed for all 56 NTM isolates what were recovered from 32 (42.1%) positive water samples, using conventional phenotypic methods, hsp65-PRA, partial sequencing of 16S rRNA and sequencing of hsp 65 genes. Results: Phenotypic evaluation and hsp65-PRA were concordant with 23 (41.1%) isolates. Also, the PRA was concordant with 16 (28.6%) and 27 (48.2%) isolates, with the partial sequencing of 16S rRNA and sequencing of hsp 65 genes, respectively. It is considered that the 19.6% (n = 11) could not be identified. Conclusion: Identification of NTM environmental isolates to the species level, especially when they are pigmented and fast-growing, both the analysis of the restriction patterns obtained by PRA and the sequencing of the 16S rRNA and hsp 65 genes are insufficient by themselves. Although they are demanding and time-consuming, biochemical tests are very useful to support data obtained by molecular methods. Instituto de Biotecnología Fil: Tortone, Claudia Andrea. Universidad Nacional de La Pampa. Facultad de Ciencias Veterinarias; Argentina Fil: Zumarraga, Martin Jose. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Gioffre, Andrea Karina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Oriani, Delia Susana. Universidad Nacional de La Pampa. Facultad de Ciencias Veterinarias; Argentina 2018-10-18T12:48:22Z 2018-10-18T12:48:22Z 2018-03 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://www.ijmyco.org/article.asp?issn=2212-5531;year=2018;volume=7;issue=1;spage=53;epage=60;aulast=Tortone http://hdl.handle.net/20.500.12123/3625 2212-5531 DOI: 10.4103/ijmy.ijmy_192_17 eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Wolters Kluwer - Medknow International Journal of Mycobacteriology 7 (1): 53-60 (Marzo 2018) |
| spellingShingle | Enzymatic Analysis Waterborne Diseases Análisis Enzimático Enfermedades Transmitidas por Agua Nontuberculous Mycobacteria Phenotypic Methods Water Sources Micobacterias no Tuberculosas Métodos Fenotípicos Fuentes de Agua Tortone, Claudia Andrea Zumarraga, Martin Jose Gioffre, Andrea Oriani, Delia Susana Utilization of molecular and conventional methods for the identification of nontuberculous mycobacteria isolated from different water sources |
| title | Utilization of molecular and conventional methods for the identification of nontuberculous mycobacteria isolated from different water sources |
| title_full | Utilization of molecular and conventional methods for the identification of nontuberculous mycobacteria isolated from different water sources |
| title_fullStr | Utilization of molecular and conventional methods for the identification of nontuberculous mycobacteria isolated from different water sources |
| title_full_unstemmed | Utilization of molecular and conventional methods for the identification of nontuberculous mycobacteria isolated from different water sources |
| title_short | Utilization of molecular and conventional methods for the identification of nontuberculous mycobacteria isolated from different water sources |
| title_sort | utilization of molecular and conventional methods for the identification of nontuberculous mycobacteria isolated from different water sources |
| topic | Enzymatic Analysis Waterborne Diseases Análisis Enzimático Enfermedades Transmitidas por Agua Nontuberculous Mycobacteria Phenotypic Methods Water Sources Micobacterias no Tuberculosas Métodos Fenotípicos Fuentes de Agua |
| url | http://www.ijmyco.org/article.asp?issn=2212-5531;year=2018;volume=7;issue=1;spage=53;epage=60;aulast=Tortone http://hdl.handle.net/20.500.12123/3625 |
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