Dados do Trabalho

Título

In vitro inactivation of SARS-CoV-2 by DMSO

Introdução

This study aimed to enhance accessibility in SARS-CoV-2 studies by investigating an integrated inactivation method within a standard drug-screening assay, eliminating the need for a separate procedure to render the virus non-infectious.

Objetivo (s)

This study investigated the use of dimethylsulfoxide (DMSO) at concentrations commonly employed in MTT assays for the inactivation of SARS-CoV-2, allowing for research to be conducted in lower biosafety level laboratories.

Material e Métodos

The experiments were conducted in a biosafety level-3 laboratory using Vero E6 cells infected with SARS-CoV-2. DMSO at concentrations of 100%, 90%, 70%, and 50% was applied to infected cells for incubation times of 15, 30, and 45 minutes. Viral viability assays were performed to assess the effectiveness of DMSO inactivation.

Resultados e Conclusão

DMSO concentrations of 100%, 90%, and 70% effectively inactivated the virus after 15 minutes of incubation. However, a 50% concentration of DMSO reduced the viral load but did not completely eradicate it. Increasing the incubation time from 15 to 45 minutes improved the inactivation efficiency. These findings suggest that DMSO inactivation of SARS-CoV-2 is concentration and time-dependent. Concentrations greater than 70% can reliably inactivate the virus in infected cells within 15 minutes. Lower concentrations on incubation times longer than 45 minutes are required for complete inactivation. The use of DMSO inactivation enables safe transfer of infected culture plates to lower containment laboratories for processing and analysis, facilitating drug screening and repurposing studies against SARS-CoV-2. It is important to note that variations in experimental conditions should be carefully evaluated, and the transfer of samples to lower containment should be performed with caution, considering the limitations of the plaque assay and the specific protocols used. These results contribute to the development of expedited research methodologies for studying SARS-CoV-2 and may aid laboratories relying on MTT assays in drug-screening and repurposing studies against SARS-CoV-2 infection.

Palavras-chave

Inactivation; DMSO; SARS-CoV-2; COVID-19; BSL-3; MTT

Agradecimentos

This study was financed in part by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS) and Banco Nacional de Desenvolvimento Econômico e Social (BNDES). Fellowship from CAPES is also acknowledged.