π Visual detection of turkey coronavirus RNA in tissues and feces by reverse-transcription loop-mediated isothermal amplification (RT-LAMP) with hydroxynaphthol blue dye
A sensitive reverse-transcription loop-mediated isothermal amplification (RT-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 Β°C for 45 min, with hydroxynaphthol blue (HNB) dye added prior to amplification. The detection limit of the RT-LAMP assay was approximately 102 EID50/50 ΞΌl 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 RT-LAMP assay. All individual feces samples collected in the field were considered positive by both conventional RT-PCR and RT-LAMP. In conclusion, RT-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. Β© 2010 Elsevier Ltd.
keywords
π samples collected (74)
π isothermal amplification (14)
π turkey coronavirus (48)
author
π€ Cardoso, Tereza C.
π€ Ferrari, Heitor F.
π€ Bregano, LΓvia C.
π€ Silva-Frade, Camila
π€ Rosa, Ana Carolina G.
π€ Andrade, Alexandre L.
year
β° 2010
issn
π 08908508
volume
24
number
6
page
415-417
citedbycount
36
download
π [BibTeX]