Detection of Mycobacterium bovis in Bovine and Bubaline tissues using Nested-PCR for TbD1
- Autores
- Araújo, Cristina Pires de; Osório, Ana Luiza Alves Rosa; Jorge, Klaudia dos Santos Goncalves; Ramos, Carlos Alberto N.; Filho, Antonio Francisco S.; Vidal, Carlos Eugênio Soto; Roxo, Eliana; Nishibe, Christiane; Almeida Junior, Nalvo Franco de; Júnior, Antônio A. F.; Silva, Marcio Roberto; Neto, Jose Diomedes Barbosa; Cerqueira, Valíria Duarte; Zumarraga, Martin Jose; Araujo, Flabio Ribeiro de
- Año de publicación
- 2014
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- In the present study, a nested-PCR system, targeting the TbD1 region, involving the performance of conventional PCR followed by real-time PCR, was developed to detect Mycobacterium bovis in bovine/bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. In terms of analytical sensitivity, the DNA of M. bovis AN5 was detected up to 1.56 ng with conventional PCR, 97.6 pg with real-time PCR, and 1.53 pg with nested-PCR in the reaction mixture. The nested-PCR exhibited 100% analytical specificity for M. bovis when tested with the DNA of reference strains of environmental mycobacteria and closely-related Actinomycetales. A clinical sensitivity value of 76.0% was detected with tissue samples from animals that exhibited positive results in the comparative intradermal tuberculin test (CITT), as well as from those with lesions compatible with tuberculosis (LCT) that rendered positive cultures. A clinical specificity value of 100% was detected with tissue samples from animals with CITT- results, with no visible lesions (NVL) and negative cultures. No significant differences were found between the nested-PCR and culture in terms of detecting CITT+ animals with LCT or with NVL. No significant differences were recorded in the detection of CITT- animals with NVL. However, nested-PCR detected a significantly higher number of positive animals than the culture in the group of animals exhibiting LCT with no previous records of CITT. The use of the nested-PCR assay to detect M. bovis in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.
Instituto de Biotecnología
Fil: Araújo, Cristina Pires de. Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-graduação em Ciência Animal; Brasil
Fil: Osório, Ana Luiza Alves Rosa. Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-graduação em Ciência Animal; Brasil
Fil: Jorge, Klaudia dos Santos Goncalves. Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-graduação em Ciência Animal; Brasil
Fil: Ramos, Carlos Alberto N. Bolsista DTI, CNPq; Brasil
Fil: Filho, Antonio Francisco S. Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-graduação em Ciência Animal; Brasil
Fil: Vidal, Carlos Eugênio Soto. Universidade Federal de Santa Maria. Programa de Pos-Graduación en Medicina Veterinaria; Brasil
Fil: Roxo, Eliana. Instituto Biológico de São Paulo; Brasil
Fil: Nishibe, Christiane. Universidade Federal de Mato Grosso do Sul. Faculdade de Computação; Brasil
Fil: Almeida Junior, Nalvo Franco de. Universidade Federal de Mato Grosso do Sul. Faculdade de Computação; Brasil
Fil: Júnior, Antônio A. F. Laboratório Nacional Agropecuário, Pedro Leopoldo; Brasil
Fil: Silva, Marcio Roberto. Empresa Brasileira de Pesquisa Agropecuária (Embrapa). Gado de Leite; Brasil
Fil: Neto, Jose Diomedes Barbosa. Universidade Federal do Pará; Brasil
Fil: Cerqueira, Valíria Duarte. Universidade Federal do Pará; Brasil
Fil: Zumarraga, Martin Jose. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Araujo, Flabio Ribeiro de. Empresa Brasileira de Pesquisa Agropecuária (Embrapa). Gado de Corte; Brasil - Fuente
- PLoS ONE 9 (3) : e91023. (2014)
- Materia
-
Mycobacterium Bovis
Tejidos Animales
Ganado Bovino
ADN
Tuberculosis
PCR
Animal Tissues
Cattle
DNA
TbD1
Polymerase Chain Reaction - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- http://creativecommons.org/licenses/by-nc-sa/4.0/
- Repositorio
.jpg)
- Institución
- Instituto Nacional de Tecnología Agropecuaria
- OAI Identificador
- oai:localhost:20.500.12123/4855
Ver los metadatos del registro completo
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Detection of Mycobacterium bovis in Bovine and Bubaline tissues using Nested-PCR for TbD1Araújo, Cristina Pires deOsório, Ana Luiza Alves RosaJorge, Klaudia dos Santos GoncalvesRamos, Carlos Alberto N.Filho, Antonio Francisco S.Vidal, Carlos Eugênio SotoRoxo, ElianaNishibe, ChristianeAlmeida Junior, Nalvo Franco deJúnior, Antônio A. F.Silva, Marcio RobertoNeto, Jose Diomedes BarbosaCerqueira, Valíria DuarteZumarraga, Martin JoseAraujo, Flabio Ribeiro deMycobacterium BovisTejidos AnimalesGanado BovinoADNTuberculosisPCRAnimal TissuesCattleDNATbD1Polymerase Chain ReactionIn the present study, a nested-PCR system, targeting the TbD1 region, involving the performance of conventional PCR followed by real-time PCR, was developed to detect Mycobacterium bovis in bovine/bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. In terms of analytical sensitivity, the DNA of M. bovis AN5 was detected up to 1.56 ng with conventional PCR, 97.6 pg with real-time PCR, and 1.53 pg with nested-PCR in the reaction mixture. The nested-PCR exhibited 100% analytical specificity for M. bovis when tested with the DNA of reference strains of environmental mycobacteria and closely-related Actinomycetales. A clinical sensitivity value of 76.0% was detected with tissue samples from animals that exhibited positive results in the comparative intradermal tuberculin test (CITT), as well as from those with lesions compatible with tuberculosis (LCT) that rendered positive cultures. A clinical specificity value of 100% was detected with tissue samples from animals with CITT- results, with no visible lesions (NVL) and negative cultures. No significant differences were found between the nested-PCR and culture in terms of detecting CITT+ animals with LCT or with NVL. No significant differences were recorded in the detection of CITT- animals with NVL. However, nested-PCR detected a significantly higher number of positive animals than the culture in the group of animals exhibiting LCT with no previous records of CITT. The use of the nested-PCR assay to detect M. bovis in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.Instituto de BiotecnologíaFil: Araújo, Cristina Pires de. Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-graduação em Ciência Animal; BrasilFil: Osório, Ana Luiza Alves Rosa. Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-graduação em Ciência Animal; BrasilFil: Jorge, Klaudia dos Santos Goncalves. Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-graduação em Ciência Animal; BrasilFil: Ramos, Carlos Alberto N. Bolsista DTI, CNPq; BrasilFil: Filho, Antonio Francisco S. Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-graduação em Ciência Animal; BrasilFil: Vidal, Carlos Eugênio Soto. Universidade Federal de Santa Maria. Programa de Pos-Graduación en Medicina Veterinaria; BrasilFil: Roxo, Eliana. Instituto Biológico de São Paulo; BrasilFil: Nishibe, Christiane. Universidade Federal de Mato Grosso do Sul. Faculdade de Computação; BrasilFil: Almeida Junior, Nalvo Franco de. Universidade Federal de Mato Grosso do Sul. Faculdade de Computação; BrasilFil: Júnior, Antônio A. F. Laboratório Nacional Agropecuário, Pedro Leopoldo; BrasilFil: Silva, Marcio Roberto. Empresa Brasileira de Pesquisa Agropecuária (Embrapa). Gado de Leite; BrasilFil: Neto, Jose Diomedes Barbosa. Universidade Federal do Pará; BrasilFil: Cerqueira, Valíria Duarte. Universidade Federal do Pará; BrasilFil: Zumarraga, Martin Jose. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Araujo, Flabio Ribeiro de. Empresa Brasileira de Pesquisa Agropecuária (Embrapa). Gado de Corte; BrasilPLOS2019-04-09T13:15:57Z2019-04-09T13:15:57Z2014-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/4855https://journals.plos.org/plosone/article?id=10.1371/journal.pone.00910231932-6203https://doi.org/10.1371/journal.pone.0091023PLoS ONE 9 (3) : e91023. (2014)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2025-10-23T11:16:54Zoai:localhost:20.500.12123/4855instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-10-23 11:16:54.98INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse |
| dc.title.none.fl_str_mv |
Detection of Mycobacterium bovis in Bovine and Bubaline tissues using Nested-PCR for TbD1 |
| title |
Detection of Mycobacterium bovis in Bovine and Bubaline tissues using Nested-PCR for TbD1 |
| spellingShingle |
Detection of Mycobacterium bovis in Bovine and Bubaline tissues using Nested-PCR for TbD1 Araújo, Cristina Pires de Mycobacterium Bovis Tejidos Animales Ganado Bovino ADN Tuberculosis PCR Animal Tissues Cattle DNA TbD1 Polymerase Chain Reaction |
| title_short |
Detection of Mycobacterium bovis in Bovine and Bubaline tissues using Nested-PCR for TbD1 |
| title_full |
Detection of Mycobacterium bovis in Bovine and Bubaline tissues using Nested-PCR for TbD1 |
| title_fullStr |
Detection of Mycobacterium bovis in Bovine and Bubaline tissues using Nested-PCR for TbD1 |
| title_full_unstemmed |
Detection of Mycobacterium bovis in Bovine and Bubaline tissues using Nested-PCR for TbD1 |
| title_sort |
Detection of Mycobacterium bovis in Bovine and Bubaline tissues using Nested-PCR for TbD1 |
| dc.creator.none.fl_str_mv |
Araújo, Cristina Pires de Osório, Ana Luiza Alves Rosa Jorge, Klaudia dos Santos Goncalves Ramos, Carlos Alberto N. Filho, Antonio Francisco S. Vidal, Carlos Eugênio Soto Roxo, Eliana Nishibe, Christiane Almeida Junior, Nalvo Franco de Júnior, Antônio A. F. Silva, Marcio Roberto Neto, Jose Diomedes Barbosa Cerqueira, Valíria Duarte Zumarraga, Martin Jose Araujo, Flabio Ribeiro de |
| author |
Araújo, Cristina Pires de |
| author_facet |
Araújo, Cristina Pires de Osório, Ana Luiza Alves Rosa Jorge, Klaudia dos Santos Goncalves Ramos, Carlos Alberto N. Filho, Antonio Francisco S. Vidal, Carlos Eugênio Soto Roxo, Eliana Nishibe, Christiane Almeida Junior, Nalvo Franco de Júnior, Antônio A. F. Silva, Marcio Roberto Neto, Jose Diomedes Barbosa Cerqueira, Valíria Duarte Zumarraga, Martin Jose Araujo, Flabio Ribeiro de |
| author_role |
author |
| author2 |
Osório, Ana Luiza Alves Rosa Jorge, Klaudia dos Santos Goncalves Ramos, Carlos Alberto N. Filho, Antonio Francisco S. Vidal, Carlos Eugênio Soto Roxo, Eliana Nishibe, Christiane Almeida Junior, Nalvo Franco de Júnior, Antônio A. F. Silva, Marcio Roberto Neto, Jose Diomedes Barbosa Cerqueira, Valíria Duarte Zumarraga, Martin Jose Araujo, Flabio Ribeiro de |
| author2_role |
author author author author author author author author author author author author author author |
| dc.subject.none.fl_str_mv |
Mycobacterium Bovis Tejidos Animales Ganado Bovino ADN Tuberculosis PCR Animal Tissues Cattle DNA TbD1 Polymerase Chain Reaction |
| topic |
Mycobacterium Bovis Tejidos Animales Ganado Bovino ADN Tuberculosis PCR Animal Tissues Cattle DNA TbD1 Polymerase Chain Reaction |
| dc.description.none.fl_txt_mv |
In the present study, a nested-PCR system, targeting the TbD1 region, involving the performance of conventional PCR followed by real-time PCR, was developed to detect Mycobacterium bovis in bovine/bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. In terms of analytical sensitivity, the DNA of M. bovis AN5 was detected up to 1.56 ng with conventional PCR, 97.6 pg with real-time PCR, and 1.53 pg with nested-PCR in the reaction mixture. The nested-PCR exhibited 100% analytical specificity for M. bovis when tested with the DNA of reference strains of environmental mycobacteria and closely-related Actinomycetales. A clinical sensitivity value of 76.0% was detected with tissue samples from animals that exhibited positive results in the comparative intradermal tuberculin test (CITT), as well as from those with lesions compatible with tuberculosis (LCT) that rendered positive cultures. A clinical specificity value of 100% was detected with tissue samples from animals with CITT- results, with no visible lesions (NVL) and negative cultures. No significant differences were found between the nested-PCR and culture in terms of detecting CITT+ animals with LCT or with NVL. No significant differences were recorded in the detection of CITT- animals with NVL. However, nested-PCR detected a significantly higher number of positive animals than the culture in the group of animals exhibiting LCT with no previous records of CITT. The use of the nested-PCR assay to detect M. bovis in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis. Instituto de Biotecnología Fil: Araújo, Cristina Pires de. Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-graduação em Ciência Animal; Brasil Fil: Osório, Ana Luiza Alves Rosa. Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-graduação em Ciência Animal; Brasil Fil: Jorge, Klaudia dos Santos Goncalves. Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-graduação em Ciência Animal; Brasil Fil: Ramos, Carlos Alberto N. Bolsista DTI, CNPq; Brasil Fil: Filho, Antonio Francisco S. Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-graduação em Ciência Animal; Brasil Fil: Vidal, Carlos Eugênio Soto. Universidade Federal de Santa Maria. Programa de Pos-Graduación en Medicina Veterinaria; Brasil Fil: Roxo, Eliana. Instituto Biológico de São Paulo; Brasil Fil: Nishibe, Christiane. Universidade Federal de Mato Grosso do Sul. Faculdade de Computação; Brasil Fil: Almeida Junior, Nalvo Franco de. Universidade Federal de Mato Grosso do Sul. Faculdade de Computação; Brasil Fil: Júnior, Antônio A. F. Laboratório Nacional Agropecuário, Pedro Leopoldo; Brasil Fil: Silva, Marcio Roberto. Empresa Brasileira de Pesquisa Agropecuária (Embrapa). Gado de Leite; Brasil Fil: Neto, Jose Diomedes Barbosa. Universidade Federal do Pará; Brasil Fil: Cerqueira, Valíria Duarte. Universidade Federal do Pará; Brasil Fil: Zumarraga, Martin Jose. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Araujo, Flabio Ribeiro de. Empresa Brasileira de Pesquisa Agropecuária (Embrapa). Gado de Corte; Brasil |
| description |
In the present study, a nested-PCR system, targeting the TbD1 region, involving the performance of conventional PCR followed by real-time PCR, was developed to detect Mycobacterium bovis in bovine/bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. In terms of analytical sensitivity, the DNA of M. bovis AN5 was detected up to 1.56 ng with conventional PCR, 97.6 pg with real-time PCR, and 1.53 pg with nested-PCR in the reaction mixture. The nested-PCR exhibited 100% analytical specificity for M. bovis when tested with the DNA of reference strains of environmental mycobacteria and closely-related Actinomycetales. A clinical sensitivity value of 76.0% was detected with tissue samples from animals that exhibited positive results in the comparative intradermal tuberculin test (CITT), as well as from those with lesions compatible with tuberculosis (LCT) that rendered positive cultures. A clinical specificity value of 100% was detected with tissue samples from animals with CITT- results, with no visible lesions (NVL) and negative cultures. No significant differences were found between the nested-PCR and culture in terms of detecting CITT+ animals with LCT or with NVL. No significant differences were recorded in the detection of CITT- animals with NVL. However, nested-PCR detected a significantly higher number of positive animals than the culture in the group of animals exhibiting LCT with no previous records of CITT. The use of the nested-PCR assay to detect M. bovis in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis. |
| publishDate |
2014 |
| dc.date.none.fl_str_mv |
2014-03 2019-04-09T13:15:57Z 2019-04-09T13:15:57Z |
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info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://purl.org/coar/resource_type/c_6501 info:ar-repo/semantics/articulo |
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article |
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publishedVersion |
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http://hdl.handle.net/20.500.12123/4855 https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0091023 1932-6203 https://doi.org/10.1371/journal.pone.0091023 |
| url |
http://hdl.handle.net/20.500.12123/4855 https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0091023 https://doi.org/10.1371/journal.pone.0091023 |
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PLOS |
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