Validation of two real-time PCRs targeting the PE-PGRS 20 gene and the region of difference 4 for the characterization of Mycobacterium bovis isolates.
Mariana Lázaro Sales,Antônio Augusto Fonseca,Livia de Lima Orzil,Andrea Padilha de Alencar,Mikael Arrais Hodon,Marina de Azevedo Issa,P. M. Soares Filho,Marcio Roberto Silva,Andrey Pereira Lage,Marcos Bryan Heinemann +9 more
TLDR
The techniques proved to be efficient, robust, sensitive, and specific for the diagnosis of M. bovis.Abstract:
This study aimed to develop and validate real-time PCR for the diagnosis of Mycobacterium bovis isolates. Two hundred and seventy-four M. bovis isolates and 156 M. tuberculosis isolates were tested. Both qPCRs amplified all of the 274 M. bovis samples, but none of the 156 M. tuberculosis samples. The qPCR for PE-PGRS 20 had 91% efficiency and a detection limit of 0.32 ng (sensitivity and specificity for qPCR "Mbovis.100" were 99.64 and 100%, respectively). The qPCR for RD4 had 100% efficiency, and a detection limit of 4 pg (diagnostic sensitivity and specificity were 100 and 100%. The qPCR tests were performed using 4 extraction sets, 3 qPCR kits, and with a range of equipment; yet, all combinations produced similar results in a diagnosticread more
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Real-time monitoring of mycobacterium genomic DNA with target-primed rolling circle amplification by a Au nanoparticle-embedded SPR biosensor.
TL;DR: A surface plasmon resonance (SPR) DNA biosensor array based on target-primed rolling circle amplification (RCA) for isothermal and rapid detection of two pathogenic mycobacteria for clinical pathogen detection holds great promise in ultrasensitive bioassay in the future.
Journal ArticleDOI
Detection of Mycobacterium bovis in artisanal cheese in the state of Pernambuco, Brazil
Renata Duarte da Silva Cezar,Norma Lucena-Silva,Jonas de Melo Borges,Vania Lucia de Assis Santana,José Wilton Pinheiro Junior +4 more
TL;DR: The results of the present study highlight the need for improving sanitary measures during the production of artisanal cheese to prevent zoonotic tuberculosis in humans, resulting from the consumption of food contaminated with M. bovis.
Journal ArticleDOI
Comparative study of Mycobacterium bovis primary isolation methods.
Marina de Azevedo Issa,Paulo Martins Soares Filho,Antônio Augusto Fonseca Júnior,Mikael Arrais Hodon,Lílian Cristian dos Santos,Jenner Karlisson Pimenta dos Reis,Rômulo Cerqueira Leite +6 more
TL;DR: Overall, the best strategy for the primary isolation of M. bovis from Brazilian samples was the decontamination with 5% sulphuric acid (final concentration) and inoculation in Middlebrook 7H11 medium formulated with OADC supplement “B”.
Journal ArticleDOI
Molecular detection of Mycobacterium bovis in cattle herds of the state of Pernambuco, Brazil
Renata Duarte da Silva Cezar,Norma Lucena-Silva,Antonio Batista Filho,Jonas de Melo Borges,Pollyane Raysa Fernandes de Oliveira,Érica Chaves Lúcio,Maíra Arruda-Lima,Vania Lucia de Assis Santana,José Wilton Pinheiro Junior +8 more
TL;DR: M. bovis DNA was detected in one milk sample what may pose a risk to public health because raw milk is commonly consumed in Brazil and the risk factors evaluated were statistically associated with BTB.
Journal ArticleDOI
Validation of a real-time PCR assay for the molecular identification of Mycobacterium tuberculosis
Mariana Lázaro Sales,Antônio Augusto Fonseca Júnior,Lívia Orzil,Andrea Padilha de Alencar,Marcio Roberto Silva,Marina de Azevedo Issa,Paulo Martins Soares Filho,Andrey Pereira Lage,Marcos Bryan Heinemann +8 more
TL;DR: This qPCR was developed with the goal of diagnosing the bacillus M. tuberculosis in samples of bacterial suspension and the correlation between tests was perfect with Kappa index of 1.0.
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