Cargando…
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...
Autores principales: | , , , , , , |
---|---|
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 |
_version_ | 1783372542311923712 |
---|---|
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. |
format | Online Article Text |
id | pubmed-6247729 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Hindawi |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT malei developmentofaconventionalrtpcrassayforrapiddetectionofporcinedeltacoronaviruswiththesamedetectionlimitasasybrgreenbasedrealtimertpcrassay AT zengfanwen developmentofaconventionalrtpcrassayforrapiddetectionofporcinedeltacoronaviruswiththesamedetectionlimitasasybrgreenbasedrealtimertpcrassay AT huangbihong developmentofaconventionalrtpcrassayforrapiddetectionofporcinedeltacoronaviruswiththesamedetectionlimitasasybrgreenbasedrealtimertpcrassay AT congfeng developmentofaconventionalrtpcrassayforrapiddetectionofporcinedeltacoronaviruswiththesamedetectionlimitasasybrgreenbasedrealtimertpcrassay AT huangren developmentofaconventionalrtpcrassayforrapiddetectionofporcinedeltacoronaviruswiththesamedetectionlimitasasybrgreenbasedrealtimertpcrassay AT majingyun developmentofaconventionalrtpcrassayforrapiddetectionofporcinedeltacoronaviruswiththesamedetectionlimitasasybrgreenbasedrealtimertpcrassay AT guopengju developmentofaconventionalrtpcrassayforrapiddetectionofporcinedeltacoronaviruswiththesamedetectionlimitasasybrgreenbasedrealtimertpcrassay |