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Fast SARS-CoV-2 detection protocol based on RNA precipitation and RT-qPCR in nasopharyngeal swab samples

Authors :
Sierra-Torre
Pampliega O
Ortiz-Zarragoitia M
Castellanos Eb
Igor Aurrekoetxea
Basaras M
Bozal-Leorri A
Unai Galicia-Garcia
Baleriola J
Maialen Sebastian-delaCruz
Asier Benito-Vicente
Ramirez-Garcia A
Aitor Rementeria
Marín-Peña A
Daniel Marino
Ane Olazagoitia-Garmendia
Sierra A
Rueda-Alaña E
Torrano
Marcos S
Leire Aparicio-Fernandez
Nora Fernandez-Jimenez
de Nanclares Gp
Buqué X
Bilbao
Guruceaga X
Patricia Aspichueta
Itziar Gonzalez-Moro
Ugo Mayor
Mendoza Lm
Areitio M
Iraia García-Santisteban
Leire Martin-Souto
Uxue Perez-Cuesta
Ainhoa Iglesias-Ara
Bernal-Chico A
Alegre Sa
Maier Lorizate
Izortze Santin
Ainara Castellanos-Rubio
Vallejo-Rodríguez J
Teresa Fuertes-Mendizábal
Serrano-Regal Mp
Tønnesen J
Irantzu Bernales
Cavaliere F
Gaminde-Blasco A
Jon Ander Nieto-Garai
César Martín
Publication Year :
2020
Publisher :
Cold Spring Harbor Laboratory, 2020.

Abstract

The SARS-CoV-2 pandemic has evolved far more aggressively in countries lacking a robust testing strategy to identify infected individuals. Given the global demand for fast and reliable diagnosis to determine the carrier individuals, a stock-out scenario for a number of essential reagents/kits used along the diagnostic process has been foreseen by many organizations. Having identified the RNA extraction step as one of the key bottlenecks, we tested several alternatives that avoid the use of commercial kits for this step. The analysis showed that 2-propanol precipitation of the viral RNA, followed by one-step RT-qPCR results in a sensitivity and specificity comparable to that provided currently by automatized systems such as the COBAS 6800 system. Therefore, this simple protocol allows SARS-CoV-2 testing independently of commercial kit providers in a time and cost-effective manner. It can be readily implemented in research and/or diagnostic laboratories worldwide, provided that patient confidentiality and researcher safety are ensured. Scaling up the testing capabilities of hospitals and research facilities will identify larger numbers of infected individuals to paint a clear picture of the COVID-19 prevalence, a pre-requisite for informed policy decision making.

Details

Language :
English
Database :
OpenAIRE
Accession number :
edsair.doi.dedup.....b7d1004a565a76cb8d38e31b57e6f953
Full Text :
https://doi.org/10.1101/2020.04.26.20081307