Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR

Detalhes bibliográficos
Autor(a) principal: Araújo, Cristina P
Data de Publicação: 2014
Outros Autores: Osório, Ana Luiza A. R, Jorge, Klaudia S. G, Ramos, Carlos A. N, Souza Filho, Antonio F, Vidal, Carlos E. S, Vargas, Agueda P. C, Roxo, Eliana, Rocha, Adalgiza S, Suffys, Philip Noel, Fonseca Jr., Antônio A, Silva, Marcio R, Barbosa Neto, José D, Cerqueira, Valíria D, Araújo, Flábio R
Tipo de documento: Artigo
Idioma: por
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/10213
Resumo: Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-Graduação em Ciência Animal, Campo Grande, MS, Brasil
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spelling Araújo, Cristina POsório, Ana Luiza A. RJorge, Klaudia S. GRamos, Carlos A. NSouza Filho, Antonio FVidal, Carlos E. SVargas, Agueda P. CRoxo, ElianaRocha, Adalgiza SSuffys, Philip NoelFonseca Jr., Antônio ASilva, Marcio RBarbosa Neto, José DCerqueira, Valíria DAraújo, Flábio R2015-05-04T17:07:31Z2015-05-04T17:07:31Z2014ARAÚJO, Cristina P. et al. Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR. Brazilian Journal of Microbiology, v.45, n.2,p.633-640, 2014.1678-4405https://www.arca.fiocruz.br/handle/icict/10213porSociedade Brasileira de MicrobiologiaInspeção SanitáriaTuberculose BovinaBovine and bubaline tuberculosisNested-PCRReal-time PCRTissueSanitary inspectionTuberculosis BovinaInspección SanitariaCiências BiológicasInspeção SanitáriaTuberculose BovinaDirect detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCRinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleUniversidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-Graduação em Ciência Animal, Campo Grande, MS, BrasilUniversidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-Graduação em Ciência Animal, Campo Grande, MS, BrasilUniversidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-Graduação em Ciência Animal, Campo Grande, MS, BrasilUniversidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Campo Grande, MS, Brasil.Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-Graduação em Ciência Animal, Campo Grande, MS, BrasilUniversidade Federal de Santa Maria. Programa de Pós-Graduação em Medicina Veterinária. Santa Maria, RS, Brasil.Universidade Federal de Santa Maria. Programa de Pós-Graduação em Medicina Veterinária. Santa Maria, RS, BrasilInstituto Biológico de São Paulo, São Paulo, SP, Brasil.Fundação Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Laboratório Nacional Agropecuário, Pedro Leopoldo, MG, Brasil.Embrapa Gado de Leite. Juiz de Fora, MG, Brasil.Universidade Federal do Pará. Castanhal, PA, Brasil.Universidade Federal do Pará. Castanhal, PA, Brasil.Embrapa Gado de Corte. Campo Grande, MS, Brasil.Post-mortem bacterial culture and specific biochemical tests are currently performed to characterize the etiologic agent of bovine tuberculosis. Cultures take up to 90 days to develop. A diagnosis by molecular tests such as PCR can provide fast and reliable results while significantly decreasing the time of confirmation. In the present study, a nested-PCR system, targeting rv2807, with conventional PCR followed by real-time PCR, was developed to detect Mycobacterium tuberculosis complex (MTC) organisms directly from bovine and 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. Regarding the analytical sensitivity, DNA of the M. bovis AN5 strain was detected up to 1.5 pg by nested-PCR, whereas DNA of M. tuberculosis H37Rv strain was detected up to 6.1 pg. The nested-PCR system showed 100% analytical specificity for MTC when tested with DNA of reference strains of non-tuberculous mycobacteria and closely-related Actinomycetales. A clinical sensitivity level of 76.7% was detected with tissues samples positive for MTC by means of the culture and conventional PCR. A clinical specificity of 100% was detected with DNA from tissue samples of cattle with negative results in the comparative intradermal tuberculin test. These cattle exhibited no visible lesions and were negative in the culture for MTC. The use of the nested-PCR assay to detect M. tuberculosis complex in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txttext/plain1914https://www.arca.fiocruz.br/bitstream/icict/10213/1/license.txt7d48279ffeed55da8dfe2f8e81f3b81fMD51ORIGINALadalgiza_rochaetal_IOC_2014.pdfapplication/pdf1284708https://www.arca.fiocruz.br/bitstream/icict/10213/2/adalgiza_rochaetal_IOC_2014.pdfa949193d633eb8179d819f773195fe4aMD52TEXTadalgiza_rochaetal_IOC_2014.pdf.txtadalgiza_rochaetal_IOC_2014.pdf.txtExtracted texttext/plain37379https://www.arca.fiocruz.br/bitstream/icict/10213/3/adalgiza_rochaetal_IOC_2014.pdf.txt9ce3ac4696e47416913498b40d96c339MD53icict/102132022-06-24 12:17:35.265oai:www.arca.fiocruz.br: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ório InstitucionalPUBhttps://www.arca.fiocruz.br/oai/requestrepositorio.arca@fiocruz.bropendoar:21352022-06-24T15:17:35Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)false
dc.title.pt_BR.fl_str_mv Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR
title Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR
spellingShingle Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR
Araújo, Cristina P
Inspeção Sanitária
Tuberculose Bovina
Bovine and bubaline tuberculosis
Nested-PCR
Real-time PCR
Tissue
Sanitary inspection
Tuberculosis Bovina
Inspección Sanitaria
Ciências Biológicas
Inspeção Sanitária
Tuberculose Bovina
title_short Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR
title_full Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR
title_fullStr Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR
title_full_unstemmed Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR
title_sort Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR
author Araújo, Cristina P
author_facet Araújo, Cristina P
Osório, Ana Luiza A. R
Jorge, Klaudia S. G
Ramos, Carlos A. N
Souza Filho, Antonio F
Vidal, Carlos E. S
Vargas, Agueda P. C
Roxo, Eliana
Rocha, Adalgiza S
Suffys, Philip Noel
Fonseca Jr., Antônio A
Silva, Marcio R
Barbosa Neto, José D
Cerqueira, Valíria D
Araújo, Flábio R
author_role author
author2 Osório, Ana Luiza A. R
Jorge, Klaudia S. G
Ramos, Carlos A. N
Souza Filho, Antonio F
Vidal, Carlos E. S
Vargas, Agueda P. C
Roxo, Eliana
Rocha, Adalgiza S
Suffys, Philip Noel
Fonseca Jr., Antônio A
Silva, Marcio R
Barbosa Neto, José D
Cerqueira, Valíria D
Araújo, Flábio R
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Araújo, Cristina P
Osório, Ana Luiza A. R
Jorge, Klaudia S. G
Ramos, Carlos A. N
Souza Filho, Antonio F
Vidal, Carlos E. S
Vargas, Agueda P. C
Roxo, Eliana
Rocha, Adalgiza S
Suffys, Philip Noel
Fonseca Jr., Antônio A
Silva, Marcio R
Barbosa Neto, José D
Cerqueira, Valíria D
Araújo, Flábio R
dc.subject.other.pt_BR.fl_str_mv Inspeção Sanitária
Tuberculose Bovina
topic Inspeção Sanitária
Tuberculose Bovina
Bovine and bubaline tuberculosis
Nested-PCR
Real-time PCR
Tissue
Sanitary inspection
Tuberculosis Bovina
Inspección Sanitaria
Ciências Biológicas
Inspeção Sanitária
Tuberculose Bovina
dc.subject.en.pt_BR.fl_str_mv Bovine and bubaline tuberculosis
Nested-PCR
Real-time PCR
Tissue
Sanitary inspection
dc.subject.es.pt_BR.fl_str_mv Tuberculosis Bovina
Inspección Sanitaria
Ciências Biológicas
dc.subject.decs.pt_BR.fl_str_mv Inspeção Sanitária
Tuberculose Bovina
description Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-Graduação em Ciência Animal, Campo Grande, MS, Brasil
publishDate 2014
dc.date.issued.fl_str_mv 2014
dc.date.accessioned.fl_str_mv 2015-05-04T17:07:31Z
dc.date.available.fl_str_mv 2015-05-04T17:07:31Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
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status_str publishedVersion
dc.identifier.citation.fl_str_mv ARAÚJO, Cristina P. et al. Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR. Brazilian Journal of Microbiology, v.45, n.2,p.633-640, 2014.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/10213
dc.identifier.issn.pt_BR.fl_str_mv 1678-4405
identifier_str_mv ARAÚJO, Cristina P. et al. Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR. Brazilian Journal of Microbiology, v.45, n.2,p.633-640, 2014.
1678-4405
url https://www.arca.fiocruz.br/handle/icict/10213
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dc.publisher.none.fl_str_mv Sociedade Brasileira de Microbiologia
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