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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...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2017
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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 |
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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 |
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