An operator-safe and simple method for SARS-CoV-2 detection using RT-qPCR without the use of a commercial RNA extraction kit

Ulloa Soledad, Bravo Cristian, Parra Bárbara, Ramirez Eugenio, Acevedo Alejandra, Fasce Rodrigo and Fernandez Jorge

Abstract

The World Health Organization has declared a pandemic caused by the new coronavirus named SARS-CoV-2. The increasing demand for commercial kits used for automated SARS-CoV-2 RNA extraction, a key stage before the RT-qPCR diagnosis, could cause stock-shortages hampering the rapid processing of samples. Although the recommendation is to use automated methods for nucleic acids extraction, alternative methods are necessary to replace commercial and these alternatives should be so secure for the laboratory operator, like automated methods. This work describes a simple, inexpensive and operator-safe method to detect SARS-CoV-2 by RT-qPCR without the need for commercial RNA extraction kits, inactivating samples by heating in presence of a stabilizer and concentrating to obtain similar results by RT-qPCR for S, N and Orf1ab genes, in comparison with automated methods

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Fecha de publicación: 2020
Idioma: ingles
Notas: Journal Virology Method