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A multiplex RT-PCR assay for rapid and simultaneous detection of four RNA viruses in swine

A multiplex reverse transcription polymerase chain rection (mRT-PCR) was developed for simultaneous detection of four RNA viruses in swine. The conserved target sequences directed to classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine epidemic dia...

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Autores principales: Zhao, Yan, Liu, Feifei, Li, Qingmei, Wu, Mengfan, Lei, Lei, Pan, Zishu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7113762/
https://www.ncbi.nlm.nih.gov/pubmed/30951787
http://dx.doi.org/10.1016/j.jviromet.2019.04.001
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author Zhao, Yan
Liu, Feifei
Li, Qingmei
Wu, Mengfan
Lei, Lei
Pan, Zishu
author_facet Zhao, Yan
Liu, Feifei
Li, Qingmei
Wu, Mengfan
Lei, Lei
Pan, Zishu
author_sort Zhao, Yan
collection PubMed
description A multiplex reverse transcription polymerase chain rection (mRT-PCR) was developed for simultaneous detection of four RNA viruses in swine. The conserved target sequences directed to classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis coronavirus (TGEV) were selected based on alignments of genomic sequences and then specific primers were designed. The mRT-PCR assay was developed and evaluated for its specificity and sensitivity. The expected product from the single viral template was amplified by mRT-PCR and no spurious PCR amplification occurred from the genomic RNA or DNA of other pathogens. For single virus or different combinations of two viruses the detection limit of mRT-PCR was consistent with a single RT-PCR wtith 1 × 10(3) copies. For different combinations of the three viruses or four viruses, sensitivity of PEDV detection partially decreased. All of positive clinical specimens by the mRT-PCR were identically confirmed using Taqman RT-qPCR. Therefore, the mRT-PCR is a useful tool for epidemiological studies and laboratory diagnosis of single virus and/or mixed infections in swine.
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spelling pubmed-71137622020-04-02 A multiplex RT-PCR assay for rapid and simultaneous detection of four RNA viruses in swine Zhao, Yan Liu, Feifei Li, Qingmei Wu, Mengfan Lei, Lei Pan, Zishu J Virol Methods Article A multiplex reverse transcription polymerase chain rection (mRT-PCR) was developed for simultaneous detection of four RNA viruses in swine. The conserved target sequences directed to classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis coronavirus (TGEV) were selected based on alignments of genomic sequences and then specific primers were designed. The mRT-PCR assay was developed and evaluated for its specificity and sensitivity. The expected product from the single viral template was amplified by mRT-PCR and no spurious PCR amplification occurred from the genomic RNA or DNA of other pathogens. For single virus or different combinations of two viruses the detection limit of mRT-PCR was consistent with a single RT-PCR wtith 1 × 10(3) copies. For different combinations of the three viruses or four viruses, sensitivity of PEDV detection partially decreased. All of positive clinical specimens by the mRT-PCR were identically confirmed using Taqman RT-qPCR. Therefore, the mRT-PCR is a useful tool for epidemiological studies and laboratory diagnosis of single virus and/or mixed infections in swine. Elsevier B.V. 2019-07 2019-04-02 /pmc/articles/PMC7113762/ /pubmed/30951787 http://dx.doi.org/10.1016/j.jviromet.2019.04.001 Text en © 2019 Elsevier B.V. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Zhao, Yan
Liu, Feifei
Li, Qingmei
Wu, Mengfan
Lei, Lei
Pan, Zishu
A multiplex RT-PCR assay for rapid and simultaneous detection of four RNA viruses in swine
title A multiplex RT-PCR assay for rapid and simultaneous detection of four RNA viruses in swine
title_full A multiplex RT-PCR assay for rapid and simultaneous detection of four RNA viruses in swine
title_fullStr A multiplex RT-PCR assay for rapid and simultaneous detection of four RNA viruses in swine
title_full_unstemmed A multiplex RT-PCR assay for rapid and simultaneous detection of four RNA viruses in swine
title_short A multiplex RT-PCR assay for rapid and simultaneous detection of four RNA viruses in swine
title_sort multiplex rt-pcr assay for rapid and simultaneous detection of four rna viruses in swine
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7113762/
https://www.ncbi.nlm.nih.gov/pubmed/30951787
http://dx.doi.org/10.1016/j.jviromet.2019.04.001
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