Rapid and sensitive detection of Bordetella bronchiseptica by loop-mediated isothermal amplification (LAMP)
Pesqui. vet. bras
;
33(10): 1222-1226, Oct. 2013. ilus, tab
Artículo
en Inglés
| LILACS
| ID: lil-697162
ABSTRACT
Bordetella bronchiseptica causes acute and chronic respiratory infections in diverse animal species and occasionally in humans. In this study, we described the establishment of a simple, sensitive and cost-efficient loop-mediated isothermal amplification (LAMP) assay for the detection of B. bronchiseptica. A set of primers towards a 235 bp region within the flagellum gene of B. bronchiseptica was designed with online software.. The specificity of the LAMP assay was examined by using 6 porcine pathogens and 100 nasal swabs collected from healthy pigs and suspect infected pigs. The results indicated that positive reactions were confirmed for all B. bronchiseptica and no cross-reactivity was observed from other non-B. bronchiseptica. In sensitivity evaluations, the technique successfully detected a serial dilutions of extracted B. bronchiseptica DNA with a detection limit of 9 copies, which was 10 times more sensitive than that of PCR. Compared with conventional PCR, the higher sensitivity of LAMP method and no need for the complex instrumentation make this LAMP assay a promising alternative for the diagnosis of B. bronchiseptica in rural areas and developing countries where there lacks of complex laboratory services.
Texto completo:
Disponible
Índice:
LILACS (Américas)
Asunto principal:
Pruebas Genéticas
/
Bordetella bronchiseptica
/
Técnicas de Amplificación de Ácido Nucleico
/
Prueba de Laboratorio
/
Flagelos
Tipo de estudio:
Estudio diagnóstico
/
Estudio pronóstico
Idioma:
Inglés
Revista:
Pesqui. vet. bras
Asunto de la revista:
Medicina Veterinaria
Año:
2013
Tipo del documento:
Artículo
País de afiliación:
China
Institución/País de afiliación:
Sichuan Agricultural University/CN
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