Literature DB >> 25036510

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.

M L Sales1, A A Fonseca1, L Orzil1, A P Alencar1, M A Hodon1, M A Issa1, P M Soares Filho1, M R Silva2, A P Lage3, M B Heinemann4.   

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 diagnostic test, demonstrating the robustness of this method. The techniques proved to be efficient, robust, sensitive, and specific for the diagnosis of M. bovis.

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Year:  2014        PMID: 25036510     DOI: 10.4238/2014.June.18.3

Source DB:  PubMed          Journal:  Genet Mol Res        ISSN: 1676-5680


  5 in total

1.  Validation of real-time PCR technique for detection of Mycobacterium bovis and Brucella abortus in bovine raw milk.

Authors:  Débora R Mascarenhas; David Germano G Schwarz; Antônio A Fonseca Júnior; Tatiana F P Oliveira; Maria A S Moreira
Journal:  Braz J Microbiol       Date:  2020-06-22       Impact factor: 2.476

2.  Molecular detection of Mycobacterium bovis in cattle herds of the state of Pernambuco, Brazil.

Authors:  Renata Duarte da Silva Cezar; Norma Lucena-Silva; Antônio Fernando Barbosa 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
Journal:  BMC Vet Res       Date:  2016-02-20       Impact factor: 2.741

3.  Comparative study of Mycobacterium bovis primary isolation methods.

Authors:  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
Journal:  Braz J Microbiol       Date:  2016-09-30       Impact factor: 2.476

4.  Matrix Assisted Laser Desorption Ionization-Time-of-Flight mass spectrometry identification of Mycobacterium bovis in Bovinae.

Authors:  Gisele Bacanelli; Larissa C Olarte; Márcio R Silva; Rudielle A Rodrigues; Paulo A M Carneiro; John B Kaneene; Taynara N Pasquatti; Haruo Takatani; Martin J Zumárraga; Rodrigo N Etges; Flábio R Araújo; Newton V Verbisck
Journal:  J Vet Med Sci       Date:  2019-08-28       Impact factor: 1.267

5.  Validation of a real-time PCR assay for the molecular identification of Mycobacterium tuberculosis.

Authors:  Mariana L Sales; Antônio Augusto Fonseca Júnior; Lívia Orzil; Andrea Padilha Alencar; Marcio Roberto Silva; Marina Azevedo Issa; Paulo Martins Soares Filho; Andrey Pereira Lage; Marcos Bryan Heinemann
Journal:  Braz J Microbiol       Date:  2015-03-04       Impact factor: 2.476

  5 in total

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