Author | Araújo, Cristina P | |
Author | Osório, Ana Luiza A. R | |
Author | Jorge, Klaudia S. G | |
Author | Ramos, Carlos A. N | |
Author | Souza Filho, Antonio F | |
Author | Vidal, Carlos E. S | |
Author | Vargas, Agueda P. C | |
Author | Roxo, Eliana | |
Author | Rocha, Adalgiza S | |
Author | Suffys, Philip Noel | |
Author | Fonseca Jr., Antônio A | |
Author | Silva, Marcio R | |
Author | Barbosa Neto, José D | |
Author | Cerqueira, Valíria D | |
Author | Araújo, Flábio R | |
Access date | 2015-05-04T17:07:31Z | |
Available date | 2015-05-04T17:07:31Z | |
Document date | 2014 | |
Citation | 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. | pt_BR |
ISSN | 1678-4405 | pt_BR |
URI | https://www.arca.fiocruz.br/handle/icict/10213 | |
Language | por | pt_BR |
Publisher | Sociedade Brasileira de Microbiologia | pt_BR |
Rights | open access | |
Subject in Portuguese | Inspeção Sanitária | pt_BR |
Subject in Portuguese | Tuberculose Bovina | pt_BR |
Title | Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR | pt_BR |
Type | Article | |
Abstract | 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. | pt_BR |
Affilliation | Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-Graduação em Ciência Animal, Campo Grande, MS, Brasil | pt_BR |
Affilliation | Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-Graduação em Ciência Animal, Campo Grande, MS, Brasil | pt_BR |
Affilliation | Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-Graduação em Ciência Animal, Campo Grande, MS, Brasil | pt_BR |
Affilliation | Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Campo Grande, MS, Brasil. | pt_BR |
Affilliation | Universidade Federal de Mato Grosso do Sul. Faculdade de Medicina Veterinária e Zootecnia. Programa de Pós-Graduação em Ciência Animal, Campo Grande, MS, Brasil | pt_BR |
Affilliation | Universidade Federal de Santa Maria. Programa de Pós-Graduação em Medicina Veterinária. Santa Maria, RS, Brasil. | pt_BR |
Affilliation | Universidade Federal de Santa Maria. Programa de Pós-Graduação em Medicina Veterinária. Santa Maria, RS, Brasil | pt_BR |
Affilliation | Instituto Biológico de São Paulo, São Paulo, SP, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Rio de Janeiro, RJ, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Rio de Janeiro, RJ, Brasil. | pt_BR |
Affilliation | Laboratório Nacional Agropecuário, Pedro Leopoldo, MG, Brasil. | pt_BR |
Affilliation | Embrapa Gado de Leite. Juiz de Fora, MG, Brasil. | pt_BR |
Affilliation | Universidade Federal do Pará. Castanhal, PA, Brasil. | pt_BR |
Affilliation | Universidade Federal do Pará. Castanhal, PA, Brasil. | pt_BR |
Affilliation | Embrapa Gado de Corte. Campo Grande, MS, Brasil. | pt_BR |
Subject | Bovine and bubaline tuberculosis | pt_BR |
Subject | Nested-PCR | pt_BR |
Subject | Real-time PCR | pt_BR |
Subject | Tissue | pt_BR |
Subject | Sanitary inspection | pt_BR |
Subject in Spanish | Tuberculosis Bovina | pt_BR |
Subject in Spanish | Inspección Sanitaria | pt_BR |
Subject in Spanish | Ciências Biológicas | pt_BR |
DeCS | Inspeção Sanitária | pt_BR |
DeCS | Tuberculose Bovina | pt_BR |