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Título: | Rapid Selective Detection of Potentially Infectious Porcine Epidemic Diarrhea Coronavirus Exposed to Heat Treatments Using Viability RT-qPCR |
Autor: | Puente, Héctor; Randazzo, Walter CSIC ORCID; Carvajal, Ana CSIC ORCID; Sánchez Moragas, Gloria CSIC ORCID | Palabras clave: | Coronavirus Porcine epidemic diarrhea virus Viability RT-qPCR Infectivity Thermal inactivation |
Fecha de publicación: | 21-ago-2020 | Editor: | Frontiers Media | Citación: | Frontiers in Microbiology 11: 1911 (2020) | Resumen: | 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. | Versión del editor: | https://doi.org/10.3389/fmicb.2020.01911 | URI: | http://hdl.handle.net/10261/218479 | DOI: | 10.3389/fmicb.2020.01911 | E-ISSN: | 1664-302X |
Aparece en las colecciones: | (IATA) Artículos (PTI Salud Global) Colección Especial COVID-19 |
Ficheros en este ítem:
Fichero | Descripción | Tamaño | Formato | |
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fmicb-11-01911.pdf | Artículo principal | 2,22 MB | Adobe PDF | Visualizar/Abrir |
FrontMicrobiol2020-Puente.pdf | Accepted manuscript | 281,46 kB | Adobe PDF | Visualizar/Abrir |
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