Title:	Detection of Mycobacterium bovis in Bovine and Bubaline Tissues Using Nested-PCR for TbD1.
Authors:	ARAUJO, C. P. OSÓRIO, A. L. A. R. JORGE, K. S. G. RAMOS, C. A. N. S. FILHO, A. F. VIDAL, C. E. S. ROXO, E. NISHIBE, c. ALMEIDA, N. F. F. JUNIOR, A. A. SILVA, M. R. B. NETO, J. D. CERQUEIRA, V. D. ZUMÁRRAGA, M. ARAUJO, F. R.
Affiliation:	MARCIO ROBERTO SILVA, CNPGL.
Date Issued:	2014
Citation:	Plos One, v. 9, n. 3, p. 1-6, 2014
Description:	In the present study, a nested-PCR system, targeting the TbD1 region, involving the performance of conventional PCR followed by real-time PCR, was developed to detect Mycobacterium bovis in bovine/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. In terms of analytical sensitivity, the DNA of M. bovis AN5 was detected up to 1.56 ng with conventional PCR, 97.6 pg with real-time PCR, and 1.53 pg with nested-PCR in the reaction mixture. The nested-PCR exhibited 100% analytical specificity for M. bovis when tested with the DNA of reference strains of environmental mycobacteria and closely-related Actinomycetales. A clinical sensitivity value of 76.0% was detected with tissue samples from animals that exhibited positive results in the comparative intradermal tuberculin test (CITT), as well as from those with lesions compatible with tuberculosis (LCT) that rendered positive cultures. A clinical specificity value of 100% was detected with tissue samples from animals with CITT- results, with no visible lesions (NVL) and negative cultures. No significant differences were found between the nested-PCR and culture in terms of detecting CITT+ animals with LCT or with NVL. No significant differences were recorded in the detection of CITT- animals with NVL. However, nested-PCR detected a significantly higher number of positive animals than the culture in the group of animals exhibiting LCT with no previous records of CITT. The use of the nested-PCR assay to detect M. bovis in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.
NAL Thesaurus:	Mycobacterium abscessus
Keywords:	Nested-PCR system Bovine and bubaline tuberculosis
Type of Material:	Artigo de periódico
Access:	openAccess
Appears in Collections:	Artigo em periódico indexado (CNPGL)