Rapid Selective Detection of Potentially Infectious Porcine Epidemic Diarrhea Coronavirus Exposed to Heat Treatments Using Viability RT-qPCR

Copyright © 2020 Puente, Randazzo, Falcó, Carvajal and Sánchez. This is an openaccess article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s...

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Autores: Puente Fernández, Héctor, Randazzo, Walter, Falcó, Irene, Carvajal Urueña, Ana María, Sánchez, Gloria
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2020
País:España
Institución:Universidad de León
Repositorio:BULERIA. Repositorio Institucional de la Universidad de León
OAI Identifier:oai:buleria.unileon.es:10612/19315
Acceso en línea:https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2020.01911/full
https://hdl.handle.net/10612/19315
Access Level:acceso abierto
Palabra clave:Sanidad animal
Coronavirus
Porcine epidemic diarrhea virus
Viability RT-qPCR
Infectivity
Thermal inactivation
3109 Ciencias Veterinarias
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spelling Rapid Selective Detection of Potentially Infectious Porcine Epidemic Diarrhea Coronavirus Exposed to Heat Treatments Using Viability RT-qPCRPuente Fernández, HéctorRandazzo, WalterFalcó, IreneCarvajal Urueña, Ana MaríaSánchez, GloriaSanidad animalCoronavirusPorcine epidemic diarrhea virusViability RT-qPCRInfectivityThermal inactivation3109 Ciencias VeterinariasCopyright © 2020 Puente, Randazzo, Falcó, Carvajal and Sánchez. This is an openaccess article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms[EN] Coronaviruses (CoVs) cause severe respiratory, enteric, and systemic infections in a wide range of hosts, including humans and animals. Porcine epidemic diarrhea virus (PEDV), a member of the Coronaviridae family, is the etiological agent of porcine epidemic diarrhea (PED), a highly contagious intestinal disease affecting pigs of all ages. In this study, we optimized a viability real-time reverse transcriptase polymerase chain reaction (RT-qPCR) for the selective detection of infectious and heat-inactivated PEDV. PEMAX™, EMA™, and PMAxx™ photoactivable dyes along with PtCl4 and CDDP platinum compounds were screened as viability markers using two RT-qPCR assays: firstly, on PEDV purified RNA, and secondly on infectious and thermally inactivated virus suspensions. Furthermore, PMAxx™ pretreatment matched the thermal inactivation pattern obtained by cell culture better than other viability markers. Finally, we further optimized the pretreatment by coupling viability markers with Triton X-100 in inoculated serum resulting in a better estimation of PEDV infectivity than RT-qPCR alone. Our study has provided a rapid analytical tool based on viability RT-qPCR to infer PEDV infectivity with potential application for feed and feed ingredients monitoring in swine industry. This development would allow for greater accuracy in epidemiological surveys and outbreak investigations.SIThis study was supported by the program of the National Institute of Agricultural and Food Research and Technology (INIA project E-RTA2015-0003-C02-02) of Spanish Government. HP was supported by FPU17/00466 predoctoral grant funded by Spanish Government. WR was supported by APOSTD/2018/150 postdoctoral grant funded by Generalitat Valenciana.Frontiers MediaSanidad AnimalFacultad de Veterinaria2020info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttps://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2020.01911/fullhttps://hdl.handle.net/10612/19315reponame:BULERIA. Repositorio Institucional de la Universidad de Leóninstname:Universidad de LeónIngléshttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:buleria.unileon.es:10612/193152026-06-24T12:43:27Z
dc.title.none.fl_str_mv Rapid Selective Detection of Potentially Infectious Porcine Epidemic Diarrhea Coronavirus Exposed to Heat Treatments Using Viability RT-qPCR
title Rapid Selective Detection of Potentially Infectious Porcine Epidemic Diarrhea Coronavirus Exposed to Heat Treatments Using Viability RT-qPCR
spellingShingle Rapid Selective Detection of Potentially Infectious Porcine Epidemic Diarrhea Coronavirus Exposed to Heat Treatments Using Viability RT-qPCR
Puente Fernández, Héctor
Sanidad animal
Coronavirus
Porcine epidemic diarrhea virus
Viability RT-qPCR
Infectivity
Thermal inactivation
3109 Ciencias Veterinarias
title_short Rapid Selective Detection of Potentially Infectious Porcine Epidemic Diarrhea Coronavirus Exposed to Heat Treatments Using Viability RT-qPCR
title_full Rapid Selective Detection of Potentially Infectious Porcine Epidemic Diarrhea Coronavirus Exposed to Heat Treatments Using Viability RT-qPCR
title_fullStr Rapid Selective Detection of Potentially Infectious Porcine Epidemic Diarrhea Coronavirus Exposed to Heat Treatments Using Viability RT-qPCR
title_full_unstemmed Rapid Selective Detection of Potentially Infectious Porcine Epidemic Diarrhea Coronavirus Exposed to Heat Treatments Using Viability RT-qPCR
title_sort Rapid Selective Detection of Potentially Infectious Porcine Epidemic Diarrhea Coronavirus Exposed to Heat Treatments Using Viability RT-qPCR
dc.creator.none.fl_str_mv Puente Fernández, Héctor
Randazzo, Walter
Falcó, Irene
Carvajal Urueña, Ana María
Sánchez, Gloria
author Puente Fernández, Héctor
author_facet Puente Fernández, Héctor
Randazzo, Walter
Falcó, Irene
Carvajal Urueña, Ana María
Sánchez, Gloria
author_role author
author2 Randazzo, Walter
Falcó, Irene
Carvajal Urueña, Ana María
Sánchez, Gloria
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Sanidad Animal
Facultad de Veterinaria
dc.subject.none.fl_str_mv Sanidad animal
Coronavirus
Porcine epidemic diarrhea virus
Viability RT-qPCR
Infectivity
Thermal inactivation
3109 Ciencias Veterinarias
topic Sanidad animal
Coronavirus
Porcine epidemic diarrhea virus
Viability RT-qPCR
Infectivity
Thermal inactivation
3109 Ciencias Veterinarias
description Copyright © 2020 Puente, Randazzo, Falcó, Carvajal and Sánchez. This is an openaccess article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms
publishDate 2020
dc.date.none.fl_str_mv 2020
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2020.01911/full
https://hdl.handle.net/10612/19315
url https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2020.01911/full
https://hdl.handle.net/10612/19315
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv http://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Frontiers Media
publisher.none.fl_str_mv Frontiers Media
dc.source.none.fl_str_mv reponame:BULERIA. Repositorio Institucional de la Universidad de León
instname:Universidad de León
instname_str Universidad de León
reponame_str BULERIA. Repositorio Institucional de la Universidad de León
collection BULERIA. Repositorio Institucional de la Universidad de León
repository.name.fl_str_mv
repository.mail.fl_str_mv
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