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Development of a Conventional RT-PCR Assay for Rapid Detection of Porcine Deltacoronavirus with the Same Detection Limit as a SYBR Green-Based Real-Time RT-PCR Assay

Porcine deltacoronavirus (PDCoV) is a newly discovered coronavirus, which belongs to the family Coronaviridae. It causes watery diarrhea, vomiting, and dehydration in newborn piglets. A sensitive RT-PCR method is urgently required to detect PDCoV infection. In this study, we developed and evaluated...

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Detalles Bibliográficos
Autores principales: Ma, Lei, Zeng, Fanwen, Huang, Bihong, Cong, Feng, Huang, Ren, Ma, Jingyun, Guo, Pengju
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6247729/
https://www.ncbi.nlm.nih.gov/pubmed/30533434
http://dx.doi.org/10.1155/2018/5035139
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author Ma, Lei
Zeng, Fanwen
Huang, Bihong
Cong, Feng
Huang, Ren
Ma, Jingyun
Guo, Pengju
author_facet Ma, Lei
Zeng, Fanwen
Huang, Bihong
Cong, Feng
Huang, Ren
Ma, Jingyun
Guo, Pengju
author_sort Ma, Lei
collection PubMed
description Porcine deltacoronavirus (PDCoV) is a newly discovered coronavirus, which belongs to the family Coronaviridae. It causes watery diarrhea, vomiting, and dehydration in newborn piglets. A sensitive RT-PCR method is urgently required to detect PDCoV infection. In this study, we developed and evaluated a conventional RT-PCR assay and a SYBR green-based real-time RT-PCR assay that targeted the PDCoV n gene. Both assays are specific and have the same limit of detection at 2 × 10(1) copies of RNA molecules per reaction. Eighty-four clinical samples were subjected to both conventional RT-PCR and real-time RT-PCR, and the same positive rate (41.7%) was achieved, which was much higher than the positive rate (26.2%) using a previously described one-step RT-PCR technique. In summary, a conventional RT-PCR technique was successfully established for the detection of PDCoV with the same detection limit as a SYBR green-based real-time RT-PCR assay.
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spelling pubmed-62477292018-12-09 Development of a Conventional RT-PCR Assay for Rapid Detection of Porcine Deltacoronavirus with the Same Detection Limit as a SYBR Green-Based Real-Time RT-PCR Assay Ma, Lei Zeng, Fanwen Huang, Bihong Cong, Feng Huang, Ren Ma, Jingyun Guo, Pengju Biomed Res Int Research Article Porcine deltacoronavirus (PDCoV) is a newly discovered coronavirus, which belongs to the family Coronaviridae. It causes watery diarrhea, vomiting, and dehydration in newborn piglets. A sensitive RT-PCR method is urgently required to detect PDCoV infection. In this study, we developed and evaluated a conventional RT-PCR assay and a SYBR green-based real-time RT-PCR assay that targeted the PDCoV n gene. Both assays are specific and have the same limit of detection at 2 × 10(1) copies of RNA molecules per reaction. Eighty-four clinical samples were subjected to both conventional RT-PCR and real-time RT-PCR, and the same positive rate (41.7%) was achieved, which was much higher than the positive rate (26.2%) using a previously described one-step RT-PCR technique. In summary, a conventional RT-PCR technique was successfully established for the detection of PDCoV with the same detection limit as a SYBR green-based real-time RT-PCR assay. Hindawi 2018-11-06 /pmc/articles/PMC6247729/ /pubmed/30533434 http://dx.doi.org/10.1155/2018/5035139 Text en Copyright © 2018 Lei Ma et al. https://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Ma, Lei
Zeng, Fanwen
Huang, Bihong
Cong, Feng
Huang, Ren
Ma, Jingyun
Guo, Pengju
Development of a Conventional RT-PCR Assay for Rapid Detection of Porcine Deltacoronavirus with the Same Detection Limit as a SYBR Green-Based Real-Time RT-PCR Assay
title Development of a Conventional RT-PCR Assay for Rapid Detection of Porcine Deltacoronavirus with the Same Detection Limit as a SYBR Green-Based Real-Time RT-PCR Assay
title_full Development of a Conventional RT-PCR Assay for Rapid Detection of Porcine Deltacoronavirus with the Same Detection Limit as a SYBR Green-Based Real-Time RT-PCR Assay
title_fullStr Development of a Conventional RT-PCR Assay for Rapid Detection of Porcine Deltacoronavirus with the Same Detection Limit as a SYBR Green-Based Real-Time RT-PCR Assay
title_full_unstemmed Development of a Conventional RT-PCR Assay for Rapid Detection of Porcine Deltacoronavirus with the Same Detection Limit as a SYBR Green-Based Real-Time RT-PCR Assay
title_short Development of a Conventional RT-PCR Assay for Rapid Detection of Porcine Deltacoronavirus with the Same Detection Limit as a SYBR Green-Based Real-Time RT-PCR Assay
title_sort development of a conventional rt-pcr assay for rapid detection of porcine deltacoronavirus with the same detection limit as a sybr green-based real-time rt-pcr assay
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6247729/
https://www.ncbi.nlm.nih.gov/pubmed/30533434
http://dx.doi.org/10.1155/2018/5035139
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