Please use this identifier to cite or link to this item: http://repositorio.ufpa.br/jspui/handle/2011/6591
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Type: Artigo de Periódico
Issue Date: Jun-2014
Authors: 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
Title: Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR
Citation: 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>.
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.
Keywords: Bovino
Bubalinos
Tuberculose em animais
Reação em cadeia da polimerase
Tecido
Inspeção sanitária
ISSN: 1517-8382
metadata.dc.rights: Acesso Aberto
Appears in Collections:Artigos Científicos - FAMEVE/CCAST

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