Abstract
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) (PtCl 4 ) 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.
Keywords: COVID-19; Fomites; SARS-CoV-2; Transmission risk; Viability RT-qPCR.
【저자키워드】 COVID-19, SARS-CoV-2, fomites, Transmission risk, viability RT-qPCR, 【초록키워드】 viruses, public health, coronavirus, pandemic, protocol, Human, Economy, risk, Spread, RNA extraction, nucleic acid, RT-qPCR, Measures, viral infectivity, nucleic acids, viability, qPCR, Cell culture, epidemiological, discrimination, Coronavirus 2019, quantification, estimation, False-positive, inactivated, PCR amplification, marker, chloride, contact with, initial assessment, viral capsid, Platinum, Suspension, SARS-CoV-2 stability, debate, PEDV, matter, false, heat, non-infectious, researcher, helping, procedure, initial, resulting, collected, develop, evaluate, reported, applied, concerning, damaged, SARS-CoV-2 particle, swabbing procedure, through contact, 【제목키워드】 non-infectious, SARS-CoV-2 particle,