Utilization of molecular and conventional methods for the identification of nontuberculous mycobacteria isolated from different water sources

Autores
Tortone, Claudia Andrea; Zumárraga, Martín José; Gioffré, Andrea Karina; Oriani, Delia Susana
Año de publicación
2018
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
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.
Fil: Tortone, Claudia Andrea. Universidad Nacional de La Pampa. Facultad de Ciencias Veterinarias; Argentina
Fil: Zumárraga, Martín José. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Gioffré, Andrea Karina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Oriani, Delia Susana. Universidad Nacional de La Pampa. Facultad de Ciencias Veterinarias; Argentina
Materia
16S RRNA SEQUENCING
HSP65 SEQUENCING
NONTUBERCULOUS MYCOBACTERIA
PHENOTYPIC METHODS
RESTRICTION ENZYME ANALYSIS-HSP65
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/178304
Acceder
id CONICETDig_2730d1f7051678c03dbca2af677c304c
oai_identifier_str oai:ri.conicet.gov.ar:11336/178304
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Utilization of molecular and conventional methods for the identification of nontuberculous mycobacteria isolated from different water sourcesTortone, Claudia AndreaZumárraga, Martín JoséGioffré, Andrea KarinaOriani, Delia Susana16S RRNA SEQUENCINGHSP65 SEQUENCINGNONTUBERCULOUS MYCOBACTERIAPHENOTYPIC METHODSRESTRICTION ENZYME ANALYSIS-HSP65https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Background: 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.Fil: Tortone, Claudia Andrea. Universidad Nacional de La Pampa. Facultad de Ciencias Veterinarias; ArgentinaFil: Zumárraga, Martín José. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaFil: Gioffré, Andrea Karina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaFil: Oriani, Delia Susana. Universidad Nacional de La Pampa. Facultad de Ciencias Veterinarias; ArgentinaWolters Kluwer Medknow Publications2018-11info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/178304Tortone, Claudia Andrea; Zumárraga, Martín José; Gioffré, Andrea Karina; Oriani, Delia Susana; Utilization of molecular and conventional methods for the identification of nontuberculous mycobacteria isolated from different water sources; Wolters Kluwer Medknow Publications; International Journal of Mycobacteriology; 7; 1; 11-2018; 53-602212-55312212-554XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.ijmyco.org/article.asp?issn=2212-5531;year=2018;volume=7;issue=1;spage=53;epage=60;aulast=Tortoneinfo:eu-repo/semantics/altIdentifier/doi/10.4103/ijmy.ijmy_192_17info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:11:15Zoai:ri.conicet.gov.ar:11336/178304instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:11:15.864CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv 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
spellingShingle Utilization of molecular and conventional methods for the identification of nontuberculous mycobacteria isolated from different water sources
Tortone, Claudia Andrea
16S RRNA SEQUENCING
HSP65 SEQUENCING
NONTUBERCULOUS MYCOBACTERIA
PHENOTYPIC METHODS
RESTRICTION ENZYME ANALYSIS-HSP65
title_short 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_sort Utilization of molecular and conventional methods for the identification of nontuberculous mycobacteria isolated from different water sources
dc.creator.none.fl_str_mv Tortone, Claudia Andrea
Zumárraga, Martín José
Gioffré, Andrea Karina
Oriani, Delia Susana
author Tortone, Claudia Andrea
author_facet Tortone, Claudia Andrea
Zumárraga, Martín José
Gioffré, Andrea Karina
Oriani, Delia Susana
author_role author
author2 Zumárraga, Martín José
Gioffré, Andrea Karina
Oriani, Delia Susana
author2_role author
author
author
dc.subject.none.fl_str_mv 16S RRNA SEQUENCING
HSP65 SEQUENCING
NONTUBERCULOUS MYCOBACTERIA
PHENOTYPIC METHODS
RESTRICTION ENZYME ANALYSIS-HSP65
topic 16S RRNA SEQUENCING
HSP65 SEQUENCING
NONTUBERCULOUS MYCOBACTERIA
PHENOTYPIC METHODS
RESTRICTION ENZYME ANALYSIS-HSP65
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv 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.
Fil: Tortone, Claudia Andrea. Universidad Nacional de La Pampa. Facultad de Ciencias Veterinarias; Argentina
Fil: Zumárraga, Martín José. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Gioffré, Andrea Karina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Oriani, Delia Susana. Universidad Nacional de La Pampa. Facultad de Ciencias Veterinarias; Argentina
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.
publishDate 2018
dc.date.none.fl_str_mv 2018-11
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/178304
Tortone, Claudia Andrea; Zumárraga, Martín José; Gioffré, Andrea Karina; Oriani, Delia Susana; Utilization of molecular and conventional methods for the identification of nontuberculous mycobacteria isolated from different water sources; Wolters Kluwer Medknow Publications; International Journal of Mycobacteriology; 7; 1; 11-2018; 53-60
2212-5531
2212-554X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/178304
identifier_str_mv Tortone, Claudia Andrea; Zumárraga, Martín José; Gioffré, Andrea Karina; Oriani, Delia Susana; Utilization of molecular and conventional methods for the identification of nontuberculous mycobacteria isolated from different water sources; Wolters Kluwer Medknow Publications; International Journal of Mycobacteriology; 7; 1; 11-2018; 53-60
2212-5531
2212-554X
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.ijmyco.org/article.asp?issn=2212-5531;year=2018;volume=7;issue=1;spage=53;epage=60;aulast=Tortone
info:eu-repo/semantics/altIdentifier/doi/10.4103/ijmy.ijmy_192_17
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Wolters Kluwer Medknow Publications
publisher.none.fl_str_mv Wolters Kluwer Medknow Publications
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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