Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR
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...
| Autor principal: | ARAÚJO, Cristina Pires de |
|---|---|
| Outros Autores: | OSÓRIO, Ana Luiza Alves Rosa, JORGE, Klaudia dos Santos Gonçalves, RAMOS, Carlos Alberto do Nascimento, SOUZA FILHO, Antonio Francisco de, VIDAL, Carlos Eugênio Soto, VARGAS, Agueda Palmira Castagna de, ROXO, Eliana, ROCHA, Adalgiza da Silva, SUFFYS, Philip Noel, FONSECA JÚNIOR, Antônio Augusto, SILVA, Marcio Roberto, BARBOSA NETO, José Diomedes, CERQUEIRA, Valíria Duarte, ARAÚJO, Flábio Ribeiro de |
| Grau: | Artigo |
| Idioma: | eng |
| Publicado em: |
2015
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| Acesso em linha: |
http://repositorio.ufpa.br/jspui/handle/2011/6591 |
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ir-2011-6591 |
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ir-2011-65912017-12-20T15:30:50Z Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR ARAÚJO, Cristina Pires de OSÓRIO, Ana Luiza Alves Rosa JORGE, Klaudia dos Santos Gonçalves RAMOS, Carlos Alberto do Nascimento SOUZA FILHO, Antonio Francisco de VIDAL, Carlos Eugênio Soto VARGAS, Agueda Palmira Castagna de ROXO, Eliana ROCHA, Adalgiza da Silva SUFFYS, Philip Noel FONSECA JÚNIOR, Antônio Augusto SILVA, Marcio Roberto BARBOSA NETO, José Diomedes CERQUEIRA, Valíria Duarte ARAÚJO, Flábio Ribeiro de Bovino Bubalinos Tuberculose em animais Reação em cadeia da polimerase Tecido Inspeção sanitária 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. 2015-04-28T17:03:40Z 2015-04-28T17:03:40Z 2014-06 Artigo de Periódico ARAUJO, Cristina P. et al. Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR. Brazilian Journal of Microbiology, São Paulo, v. 45, n. 2, p. 633-640, abr./jun. 2014. Disponível em: <http://www.scielo.br/pdf/bjm/v45n2/35.pdf>. Acesso em: 24 abr. 2015. <http://dx.doi.org/10.1590/S1517-83822014000200035>. 1517-8382 http://repositorio.ufpa.br/jspui/handle/2011/6591 eng Acesso Aberto application/pdf |
| institution |
Repositório Institucional - Universidade Federal do Pará |
| collection |
RI-UFPA |
| language |
eng |
| topic |
Bovino Bubalinos Tuberculose em animais Reação em cadeia da polimerase Tecido Inspeção sanitária |
| spellingShingle |
Bovino Bubalinos Tuberculose em animais Reação em cadeia da polimerase Tecido Inspeção sanitária ARAÚJO, Cristina Pires de Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR |
| topic_facet |
Bovino Bubalinos Tuberculose em animais Reação em cadeia da polimerase Tecido Inspeção sanitária |
| description |
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. |
| format |
Artigo |
| author |
ARAÚJO, Cristina Pires de |
| author2 |
OSÓRIO, Ana Luiza Alves Rosa JORGE, Klaudia dos Santos Gonçalves RAMOS, Carlos Alberto do Nascimento SOUZA FILHO, Antonio Francisco de VIDAL, Carlos Eugênio Soto VARGAS, Agueda Palmira Castagna de ROXO, Eliana ROCHA, Adalgiza da Silva SUFFYS, Philip Noel FONSECA JÚNIOR, Antônio Augusto SILVA, Marcio Roberto BARBOSA NETO, José Diomedes CERQUEIRA, Valíria Duarte ARAÚJO, Flábio Ribeiro de |
| author2Str |
OSÓRIO, Ana Luiza Alves Rosa JORGE, Klaudia dos Santos Gonçalves RAMOS, Carlos Alberto do Nascimento SOUZA FILHO, Antonio Francisco de VIDAL, Carlos Eugênio Soto VARGAS, Agueda Palmira Castagna de ROXO, Eliana ROCHA, Adalgiza da Silva SUFFYS, Philip Noel FONSECA JÚNIOR, Antônio Augusto SILVA, Marcio Roberto BARBOSA NETO, José Diomedes CERQUEIRA, Valíria Duarte ARAÚJO, Flábio Ribeiro de |
| title |
Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR |
| 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 |
| publishDate |
2015 |
| url |
http://repositorio.ufpa.br/jspui/handle/2011/6591 |
| _version_ |
1832602560303726592 |
| score |
11.755432 |