Platinum chloride-based viability RT-qPCR for SARS-CoV-2 detection in complex samples

Abstract Isolation, contact tracing and restrictions on social movement are being globally implemented to prevent and control onward spread of SARS-CoV-2, even though the infection risk modelled on RNA detection by RT-qPCR remains biased as viral shedding and infectivity are not discerned. Thus, we...

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Autores principales:	Enric Cuevas-Ferrando, Walter Randazzo, Alba Pérez-Cataluña, Irene Falcó, David Navarro, Sandra Martin-Latil, Azahara Díaz-Reolid, Inés Girón-Guzmán, Ana Allende, Gloria Sánchez
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/40477fffc02543c594f4fe9a55467df4
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spelling	oai:doaj.org-article:40477fffc02543c594f4fe9a55467df42021-12-02T18:02:23ZPlatinum chloride-based viability RT-qPCR for SARS-CoV-2 detection in complex samples10.1038/s41598-021-97700-x2045-2322https://doaj.org/article/40477fffc02543c594f4fe9a55467df42021-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-97700-xhttps://doaj.org/toc/2045-2322Abstract Isolation, contact tracing and restrictions on social movement are being globally implemented to prevent and control onward spread of SARS-CoV-2, even though the infection risk modelled on RNA detection by RT-qPCR remains biased as viral shedding and infectivity are not discerned. Thus, we aimed to develop a rapid viability RT-qPCR procedure to infer SARS-CoV-2 infectivity in clinical specimens and environmental samples. We screened monoazide dyes and platinum compounds as viability molecular markers on five SARS-CoV-2 RNA targets. A platinum chloride-based viability RT-qPCR was then optimized using genomic RNA, and inactivated SARS-CoV-2 particles inoculated in buffer, stool, and urine. Our results were finally validated in nasopharyngeal swabs from persons who tested positive for COVID-19 and in wastewater samples positive for SARS-CoV-2 RNA. We established a rapid viability RT-qPCR that selectively detects potentially infectious SARS-CoV-2 particles in complex matrices. In particular, the confirmed positivity of nasopharyngeal swabs following the viability procedure suggests their potential infectivity, while the complete prevention of amplification in wastewater indicated either non-infectious particles or free RNA. The viability RT-qPCR approach provides a more accurate ascertainment of the infectious viruses detection and it may complement analyses to foster risk-based investigations for the prevention and control of new or re-occurring outbreaks with a broad application spectrum.Enric Cuevas-FerrandoWalter RandazzoAlba Pérez-CataluñaIrene FalcóDavid NavarroSandra Martin-LatilAzahara Díaz-ReolidInés Girón-GuzmánAna AllendeGloria SánchezNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Enric Cuevas-Ferrando Walter Randazzo Alba Pérez-Cataluña Irene Falcó David Navarro Sandra Martin-Latil Azahara Díaz-Reolid Inés Girón-Guzmán Ana Allende Gloria Sánchez Platinum chloride-based viability RT-qPCR for SARS-CoV-2 detection in complex samples
description	Abstract Isolation, contact tracing and restrictions on social movement are being globally implemented to prevent and control onward spread of SARS-CoV-2, even though the infection risk modelled on RNA detection by RT-qPCR remains biased as viral shedding and infectivity are not discerned. Thus, we aimed to develop a rapid viability RT-qPCR procedure to infer SARS-CoV-2 infectivity in clinical specimens and environmental samples. We screened monoazide dyes and platinum compounds as viability molecular markers on five SARS-CoV-2 RNA targets. A platinum chloride-based viability RT-qPCR was then optimized using genomic RNA, and inactivated SARS-CoV-2 particles inoculated in buffer, stool, and urine. Our results were finally validated in nasopharyngeal swabs from persons who tested positive for COVID-19 and in wastewater samples positive for SARS-CoV-2 RNA. We established a rapid viability RT-qPCR that selectively detects potentially infectious SARS-CoV-2 particles in complex matrices. In particular, the confirmed positivity of nasopharyngeal swabs following the viability procedure suggests their potential infectivity, while the complete prevention of amplification in wastewater indicated either non-infectious particles or free RNA. The viability RT-qPCR approach provides a more accurate ascertainment of the infectious viruses detection and it may complement analyses to foster risk-based investigations for the prevention and control of new or re-occurring outbreaks with a broad application spectrum.
format	article
author	Enric Cuevas-Ferrando Walter Randazzo Alba Pérez-Cataluña Irene Falcó David Navarro Sandra Martin-Latil Azahara Díaz-Reolid Inés Girón-Guzmán Ana Allende Gloria Sánchez
author_facet	Enric Cuevas-Ferrando Walter Randazzo Alba Pérez-Cataluña Irene Falcó David Navarro Sandra Martin-Latil Azahara Díaz-Reolid Inés Girón-Guzmán Ana Allende Gloria Sánchez
author_sort	Enric Cuevas-Ferrando
title	Platinum chloride-based viability RT-qPCR for SARS-CoV-2 detection in complex samples
title_short	Platinum chloride-based viability RT-qPCR for SARS-CoV-2 detection in complex samples
title_full	Platinum chloride-based viability RT-qPCR for SARS-CoV-2 detection in complex samples
title_fullStr	Platinum chloride-based viability RT-qPCR for SARS-CoV-2 detection in complex samples
title_full_unstemmed	Platinum chloride-based viability RT-qPCR for SARS-CoV-2 detection in complex samples
title_sort	platinum chloride-based viability rt-qpcr for sars-cov-2 detection in complex samples
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/40477fffc02543c594f4fe9a55467df4
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Platinum chloride-based viability RT-qPCR for SARS-CoV-2 detection in complex samples

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