Discrimination of non-infectious SARS-CoV-2 particles from fomites by viability RT-qPCR

Autor: Walter Randazzo, Gloria Sánchez, Rosa Aznar, Azahara Díaz-Reolid, Inés Girón-Guzmán, Irene Falcó, Enric Cuevas-Ferrando, Alba Pérez-Cataluña
Přispěvatelé: Generalitat Valenciana, Consejo Superior de Investigaciones Científicas (España), Ministerio de Ciencia e Innovación (España), European Commission, EIT Food
Rok vydání: 2021
Předmět:
Zdroj: Environmental Research
Digital.CSIC: Repositorio Institucional del CSIC
Consejo Superior de Investigaciones Científicas (CSIC)
Digital.CSIC. Repositorio Institucional del CSIC
instname
ISSN: 1096-0953
Popis: The ongoing coronavirus 2019 (COVID-19) pandemic constitutes a concerning global threat to public health and economy. In the midst of this pandemic scenario, the role of environment-to-human COVID-19 spread is still a matter of debate because mixed results have been reported concerning SARS-CoV-2 stability on high-touch surfaces in real-life scenarios. Up to now, no alternative and accessible procedures for cell culture have been applied to evaluate SARS-CoV-2 infectivity on fomites. Several strategies based on viral capsid integrity have latterly been developed using viability markers to selectively remove false-positive qPCR signals resulting from free nucleic acids and damaged viruses. These have finally allowed an estimation of viral infectivity. The present study aims to provide a rapid molecular-based protocol for detection and quantification of viable SARS-CoV-2 from fomites based on the discrimination of non-infectious SARS-CoV-2 particles by platinum chloride (IV) (PtCl4) viability RT-qPCR. An initial assessment compared two different swabbing procedures to recover inactivated SARS-CoV-2 particles from fomites coupled with two RNA extraction methods. Procedures were validated with human (E229) and porcine (PEDV) coronavirus surrogates, and compared with inactivated SARS-CoV-2 suspensions on glass, steel and plastic surfaces. The viability RT-qPCR efficiently removed the PCR amplification signals from heat and gamma-irradiated inactivated SARS-CoV-2 suspensions that had been collected from specified surfaces. This study proposes a rapid viability RT-qPCR that discriminates non-infectious SARS-CoV-2 particles on surfaces thus helping researchers to better understand the risk of contracting COVID-19 through contact with fomites and to develop more efficient epidemiological measures.
The study was supported by EIT-Food (COVID-19 BAEMitup), CSIC (202070E101), Generalitat Valenciana (Covid_19-SCI), MICINN co-founded by AEI/FEDER, UE (AGL2017-82909), and MICINN/AEI (PID2019-105509RJ-I00). EC-F is recipient of a predoctoral contract from the MICINN, Call 2018. IGG is recipient of a Collaboration Grant 2020–2021 from the MEFP.
Databáze: OpenAIRE
Externí odkaz: