A simple and rapid identification method for newly emerged porcine Deltacoronavirus with loop-mediated isothermal amplification

A simple and rapid identification method for newly emerged porcine Deltacoronavirus with loop-mediated isothermal amplification

Abstract Background Porcine Deltacoronavirus (PDCoV) is a newly emerged enteropathogenic coronavirus that causes diarrhea and mortality in neonatal piglets. PDCoV has spread to many countries around the world, leading to significant economic losses in the pork industry. Therefore, a rapid and sens...

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Autores principales: Zhang,Fanfan, Ye,Yu, Song,Deping, Guo,Nannan, Peng,Qi, Li,Anqi, Zhou,Xingrong, Chen,Yanjun, Zhang,Min, Huang,Dongyan, Tang,Yuxin
Lenguaje:English
Publicado: Sociedad de Biología de Chile 2017
Materias:
Porcine Deltacoronavirus (PDCoV)
RT-LAMP
Rapid diagnosis
Acceso en línea:http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602017000100222
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spelling oai:scielo:S0716-976020170001002222017-11-03A simple and rapid identification method for newly emerged porcine Deltacoronavirus with loop-mediated isothermal amplificationZhang,FanfanYe,YuSong,DepingGuo,NannanPeng,QiLi,AnqiZhou,XingrongChen,YanjunZhang,MinHuang,DongyanTang,Yuxin Porcine Deltacoronavirus (PDCoV) RT-LAMP Rapid diagnosis Abstract Background Porcine Deltacoronavirus (PDCoV) is a newly emerged enteropathogenic coronavirus that causes diarrhea and mortality in neonatal piglets. PDCoV has spread to many countries around the world, leading to significant economic losses in the pork industry. Therefore, a rapid and sensitive method for detection of PDCoV in clinical samples is urgently needed. Results In this study, we developed a single-tube one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay specific for nucleocapsid gene to diagnose and monitor PDCoV infections. The detection limit of RT-LAMP assay was 1 × 101 copies of PDCoV, which was approximately 100-fold more sensitive than gel-based one-step reverse transcription polymerase chain reaction (RT-PCR). This assay could specifically amplify PDCoV and had no cross amplification with porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine kobuvirus (PKoV), porcine astrovirus (PAstV), porcine reproductive and respiratory syndrome virus (PRRSV), classic swine fever virus (CSFV), and porcine circovirus type 2 (PCV2). By screening a panel of clinical specimens (N = 192), this method presented a similar sensitivity with nested RT-PCR and was 1–2 log more sensitive than conventional RT-PCR in detection of PDCoV. Conclusions The RT-LAMP assay established in this study is a potentially valuable tool, especially in low-resource laboratories and filed settings, for a rapid diagnosis, surveillance, and molecular epidemiology investigation of PDCoV infections. To the best of our knowledge, this is the first work for detection of newly emerged PDCoV with LAMP technology.info:eu-repo/semantics/openAccessSociedad de Biología de ChileBiological Research v.50 20172017-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602017000100222en10.1186/s40659-017-0135-6
institution Scielo Chile
collection Scielo Chile
language English
topic Porcine Deltacoronavirus (PDCoV)
RT-LAMP
Rapid diagnosis
spellingShingle Porcine Deltacoronavirus (PDCoV)
RT-LAMP
Rapid diagnosis
Zhang,Fanfan
Ye,Yu
Song,Deping
Guo,Nannan
Peng,Qi
Li,Anqi
Zhou,Xingrong
Chen,Yanjun
Zhang,Min
Huang,Dongyan
Tang,Yuxin
A simple and rapid identification method for newly emerged porcine Deltacoronavirus with loop-mediated isothermal amplification
description Abstract Background Porcine Deltacoronavirus (PDCoV) is a newly emerged enteropathogenic coronavirus that causes diarrhea and mortality in neonatal piglets. PDCoV has spread to many countries around the world, leading to significant economic losses in the pork industry. Therefore, a rapid and sensitive method for detection of PDCoV in clinical samples is urgently needed. Results In this study, we developed a single-tube one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay specific for nucleocapsid gene to diagnose and monitor PDCoV infections. The detection limit of RT-LAMP assay was 1 × 101 copies of PDCoV, which was approximately 100-fold more sensitive than gel-based one-step reverse transcription polymerase chain reaction (RT-PCR). This assay could specifically amplify PDCoV and had no cross amplification with porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine kobuvirus (PKoV), porcine astrovirus (PAstV), porcine reproductive and respiratory syndrome virus (PRRSV), classic swine fever virus (CSFV), and porcine circovirus type 2 (PCV2). By screening a panel of clinical specimens (N = 192), this method presented a similar sensitivity with nested RT-PCR and was 1–2 log more sensitive than conventional RT-PCR in detection of PDCoV. Conclusions The RT-LAMP assay established in this study is a potentially valuable tool, especially in low-resource laboratories and filed settings, for a rapid diagnosis, surveillance, and molecular epidemiology investigation of PDCoV infections. To the best of our knowledge, this is the first work for detection of newly emerged PDCoV with LAMP technology.
author Zhang,Fanfan
Ye,Yu
Song,Deping
Guo,Nannan
Peng,Qi
Li,Anqi
Zhou,Xingrong
Chen,Yanjun
Zhang,Min
Huang,Dongyan
Tang,Yuxin
author_facet Zhang,Fanfan
Ye,Yu
Song,Deping
Guo,Nannan
Peng,Qi
Li,Anqi
Zhou,Xingrong
Chen,Yanjun
Zhang,Min
Huang,Dongyan
Tang,Yuxin
author_sort Zhang,Fanfan
title A simple and rapid identification method for newly emerged porcine Deltacoronavirus with loop-mediated isothermal amplification
title_short A simple and rapid identification method for newly emerged porcine Deltacoronavirus with loop-mediated isothermal amplification
title_full A simple and rapid identification method for newly emerged porcine Deltacoronavirus with loop-mediated isothermal amplification
title_fullStr A simple and rapid identification method for newly emerged porcine Deltacoronavirus with loop-mediated isothermal amplification
title_full_unstemmed A simple and rapid identification method for newly emerged porcine Deltacoronavirus with loop-mediated isothermal amplification
title_sort simple and rapid identification method for newly emerged porcine deltacoronavirus with loop-mediated isothermal amplification
publisher Sociedad de Biología de Chile
publishDate 2017
url http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602017000100222
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