Use of touch-down polymerase chain reaction to enhance the sensitivity of Mycobacterium bovis detection

The confirmatory diagnosis of Mycobacterium bovis (M. bovis) in animal samples is carried out by culture in Stonebrink media. However, culture is very slow because of the extremely long duplication time of the bacillus and difficult because of the scarcity of bacilli in diagnostic samples. This stud...

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Detalles Bibliográficos
Autores principales: Zumarraga, Martin Jose, Meikle, Virginia, Bernardelli, Amelia, Abdala, Alejandro Ariel, Tarabla, Hector Dante, Romano, Maria Isabel, Cataldi, Angel Adrian
Formato: info:ar-repo/semantics/artículo
Lenguaje:Inglés
Publicado: Sage Publications 2020
Materias:
Acceso en línea:http://hdl.handle.net/20.500.12123/8231
https://journals.sagepub.com/doi/10.1177/104063870501700303
https://doi.org/10.1177/104063870501700303
Descripción
Sumario:The confirmatory diagnosis of Mycobacterium bovis (M. bovis) in animal samples is carried out by culture in Stonebrink media. However, culture is very slow because of the extremely long duplication time of the bacillus and difficult because of the scarcity of bacilli in diagnostic samples. This study describes the development of a single-tube touch-down polymerase chain reaction (PCR) protocol for the detection of M. bovis using primers that target the IS6110 element. Spiked water and milk as well as routine diagnostic samples (milk and nasal swabs) from M. bovis–positive cattle were tested. This protocol allows the rapid and sensitive detection of M. bovis in bovine samples by enhancing the sensitivity of standard PCR amplification.