Visual detection of turkey coronavirus RNA in tissues and feces by reverse-transcription loop-mediated isothermal amplification (RT-LAMP) with hydroxynaphthol blue dye

被引：53

作者：

Cardoso, Tereza C. ^{[1
]}

Ferrari, Heitor F. ^{[1
]}

Bregano, Livia C. ^{[1
]}

Silva-Frade, Camila ^{[1
]}

Rosa, Ana Carolina G. ^{[1
]}

Andrade, Alexandre L. ^{[1
]}

机构：

[1] UNESP Sao Paulo State Univ, Lab Anim Virol, Sao Paulo, Brazil

来源：

MOLECULAR AND CELLULAR PROBES | 2010年 / 24卷 / 06期

基金：

巴西圣保罗研究基金会;

关键词：

Turkey coronavirus; Hydroxynaphthol blue; LAMP assay; Spike gene;

D O I：

10.1016/j.mcp.2010.08.003

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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

引用

页码：415 / 417

页数：3

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