UnCovid: A versatile, low-cost, and open-source protocol for SARS-CoV-2 RNA detection

Roberto Alcántara; Katherin Peñaranda; Gabriel Mendoza-Rojas; Jose A Nakamoto; Eva Dueñas; Daniela Alvarez; Vanessa Adaui; Pohl Milón

doi:10.1016/j.xpro.2021.100878

UnCovid: A versatile, low-cost, and open-source protocol for SARS-CoV-2 RNA detection

STAR Protoc. 2021 Dec 17;2(4):100878. doi: 10.1016/j.xpro.2021.100878. Epub 2021 Sep 25.

Authors

Roberto Alcántara¹, Katherin Peñaranda¹, Gabriel Mendoza-Rojas¹, Jose A Nakamoto¹, Eva Dueñas¹, Daniela Alvarez¹, Vanessa Adaui¹, Pohl Milón¹

Affiliation

¹ Centre for Research and Innovation, Health Sciences Faculty, Universidad Peruana de Ciencias Aplicadas (UPC), Lima 15023, Peru.

Abstract

Here, we describe a detailed step-by-step protocol to detect SARS-CoV-2 RNA using RT-PCR-mediated amplification and CRISPR/Cas-based visualization. The optimized assay uses basic molecular biology equipment such as conventional thermocyclers and transilluminators for qualitative detection. Alternatively, a fluorescence plate reader can be used for quantitative measurements. The protocol detects two regions of the SARS-CoV-2 genome in addition to the human RNaseP sample control. Aiming to reach remote regions, this work was developed to use the portable molecular workstation from BentoLab. For complete details on the use and execution of this protocol, please refer to Alcántara et al., 2021.

Keywords: Biotechnology and bioengineering; CRISPR; Clinical Protocol; Immunology; Microbiology; Molecular Biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

COVID-19 / diagnosis*
COVID-19 / genetics
COVID-19 / virology
CRISPR-Cas Systems*
Humans
Nucleic Acid Amplification Techniques / methods*
RNA, Viral / analysis*
RNA, Viral / genetics*
SARS-CoV-2 / genetics*
SARS-CoV-2 / isolation & purification

Substances

RNA, Viral