Cargando…

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

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 meth...

Descripción completa

Detalles Bibliográficos
Autores principales: Zhang, Fanfan, Ye, Yu, Song, Deping, Guo, Nannan, Peng, Qi, Li, Anqi, Zhou, Xingrong, Chen, Yanjun, Zhang, Min, Huang, Dongyan, Tang, Yuxin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5607838/
https://www.ncbi.nlm.nih.gov/pubmed/28934984
http://dx.doi.org/10.1186/s40659-017-0135-6
_version_ 1783265349724012544
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
collection PubMed
description 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 × 10(1) 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s40659-017-0135-6) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-5607838
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-56078382017-09-24 A simple and rapid identification method for newly emerged porcine Deltacoronavirus with loop-mediated isothermal amplification Zhang, Fanfan Ye, Yu Song, Deping Guo, Nannan Peng, Qi Li, Anqi Zhou, Xingrong Chen, Yanjun Zhang, Min Huang, Dongyan Tang, Yuxin Biol Res Research Article 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 × 10(1) 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s40659-017-0135-6) contains supplementary material, which is available to authorized users. BioMed Central 2017-09-21 /pmc/articles/PMC5607838/ /pubmed/28934984 http://dx.doi.org/10.1186/s40659-017-0135-6 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
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
title 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_short 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
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5607838/
https://www.ncbi.nlm.nih.gov/pubmed/28934984
http://dx.doi.org/10.1186/s40659-017-0135-6
work_keys_str_mv AT zhangfanfan asimpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT yeyu asimpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT songdeping asimpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT guonannan asimpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT pengqi asimpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT lianqi asimpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT zhouxingrong asimpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT chenyanjun asimpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT zhangmin asimpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT huangdongyan asimpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT tangyuxin asimpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT zhangfanfan simpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT yeyu simpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT songdeping simpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT guonannan simpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT pengqi simpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT lianqi simpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT zhouxingrong simpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT chenyanjun simpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT zhangmin simpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT huangdongyan simpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification
AT tangyuxin simpleandrapididentificationmethodfornewlyemergedporcinedeltacoronaviruswithloopmediatedisothermalamplification