LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis
Braz. j. microbiol
; 46(2): 619-626, Apr-Jun/2015. tab, graf
Article
en En
| LILACS
| ID: lil-749730
Biblioteca responsable:
BR1.1
ABSTRACT
In this study, we developed new sets of primers to detect Brucella spp. and M. avium subsp. paratuberculosis (MAP) through isothermal amplification. We selected a previously well-characterized target gene, bscp31, specific for Brucella spp. and IS900 for MAP. The limits of detection using the loop-mediated isothermal amplification (LAMP) protocols described herein were similar to those of conventional PCR targeting the same sequences. Hydroxynaphtol blue and SYBR GreenTM allowed direct naked-eye detection with identical sensitivity as agarose gel electrophoresis. We included the LAMP-based protocol in a rapid identification scheme of the respective pathogens, and all tested isolates were correctly identified within 2 to 3 h. In addition, both protocols were suitable for specifically identifying the respective pathogens; in the case of Brucella, it also allowed the identification of all the biovars tested. We conclude that LAMP is a suitable rapid molecular typing tool that could help to shorten the time required to identify insidious bacteria in low-complexity laboratories, mainly in developing countries.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
LILACS
Asunto principal:
Brucella
/
Mycobacterium avium subsp. paratuberculosis
/
Técnicas de Amplificación de Ácido Nucleico
/
Técnicas de Diagnóstico Molecular
Tipo de estudio:
Diagnostic_studies
/
Evaluation_studies
/
Prognostic_studies
Límite:
Animals
/
Humans
Idioma:
En
Revista:
Braz. j. microbiol
Asunto de la revista:
MICROBIOLOGIA
Año:
2015
Tipo del documento:
Article
/
Project document
País de afiliación:
Argentina
Pais de publicación:
Brasil