Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Outros Autores: | , , , , , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Brazilian Journal of Microbiology |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000200035 |
Resumo: | 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. |
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Brazilian Journal of Microbiology |
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Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCRbovine and bubaline tuberculosisnested-PCRreal-time PCRtissuesanitary inspectionPost-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.Sociedade Brasileira de Microbiologia2014-06-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000200035Brazilian Journal of Microbiology v.45 n.2 2014reponame:Brazilian Journal of Microbiologyinstname:Sociedade Brasileira de Microbiologia (SBM)instacron:SBM10.1590/S1517-83822014000200035info:eu-repo/semantics/openAccessAraújo,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,ElianaRocha,Adalgiza S.Suffys,Philip N.Fonseca Júnior,Antônio A.Silva,Marcio R.Barbosa Neto,José D.Cerqueira,Valíria D.Araújo,Flábio R.eng2014-09-30T00:00:00Zoai:scielo:S1517-83822014000200035Revistahttps://www.scielo.br/j/bjm/ONGhttps://old.scielo.br/oai/scielo-oai.phpbjm@sbmicrobiologia.org.br||mbmartin@usp.br1678-44051517-8382opendoar:2014-09-30T00:00Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM)false |
dc.title.none.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. bovine and bubaline tuberculosis nested-PCR real-time PCR tissue sanitary inspection |
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 N. Fonseca Júnior,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 N. Fonseca Júnior,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 N. Fonseca Júnior,Antônio A. Silva,Marcio R. Barbosa Neto,José D. Cerqueira,Valíria D. Araújo,Flábio R. |
dc.subject.por.fl_str_mv |
bovine and bubaline tuberculosis nested-PCR real-time PCR tissue sanitary inspection |
topic |
bovine and bubaline tuberculosis nested-PCR real-time PCR tissue sanitary inspection |
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. |
publishDate |
2014 |
dc.date.none.fl_str_mv |
2014-06-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000200035 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000200035 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S1517-83822014000200035 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Sociedade Brasileira de Microbiologia |
publisher.none.fl_str_mv |
Sociedade Brasileira de Microbiologia |
dc.source.none.fl_str_mv |
Brazilian Journal of Microbiology v.45 n.2 2014 reponame:Brazilian Journal of Microbiology instname:Sociedade Brasileira de Microbiologia (SBM) instacron:SBM |
instname_str |
Sociedade Brasileira de Microbiologia (SBM) |
instacron_str |
SBM |
institution |
SBM |
reponame_str |
Brazilian Journal of Microbiology |
collection |
Brazilian Journal of Microbiology |
repository.name.fl_str_mv |
Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM) |
repository.mail.fl_str_mv |
bjm@sbmicrobiologia.org.br||mbmartin@usp.br |
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1752122206526111744 |