Otro
Visual detection of turkey coronavirus RNA in tissues and feces by reverse-transcription loop-mediated isothermal amplification (RT-LAMP) with hydroxynaphthol blue dye
Registro en:
Molecular and Cellular Probes. London: Academic Press Ltd- Elsevier B.V. Ltd, v. 24, n. 6, p. 415-417, 2010.
0890-8508
10.1016/j.mcp.2010.08.003
WOS:000283970200015
Autor
Cardoso, Tereza C.
Ferrari, Heitor F.
Bregano, Livia C.
Silva-Frade, Camila
Rosa, Ana Carolina G.
Andrade, Alexandre Lima de
Resumen
A sensitive reverse-transcription loop-mediated isothermal amplification (RI-LAMP) assay was developed for the rapid visual detection of turkey coronavirus (TCoV) infection The reaction is performed in one step in a single tube at 65 degrees C for 45 min with hydroxynaphthol blue (HNB) dye added prior to amplification The detection limit of the RI-LAMP assay was approximately 10(2) EID50/50 mu l TCoV genome and no cross-reaction with other avian viruses was observed The assay was evaluated further in tissue suspensions prepared from the ileum and ileum-caecal junctions of infected turkey embryos 100% of these samples were positive in the RI-LAMP assay All individual feces samples collected in the field were considered positive by both conventional RT-PCR and RI-LAMP In conclusion RI-LAMP with HNB dye was shown to be a sensitive simple assay for the rapid diagnosis of TCoV infection either directly from feces or in association with virus isolation methods (C) 2010 Elsevier Ltd All rights reserved Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)