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Development of an alternative saliva test for diagnosis of SARS-CoV-2 using TRIzol: Adapting to countries with lower incomes looking for a large-scale detection program

The use of saliva for the diagnosis of SARS-CoV-2 has shown to be a good alternative to nasopharyngeal swabs (NPS), since it permits self-collection, avoids the exposure of healthy persons to infected patients, reduces waiting times, eliminates the need of personal protective equipment and is non-in...

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Autores principales: Miranda-Ortiz, Haydee, Fernández-Figueroa, Edith A., Ruíz-García, Erika B., Muñoz-Rivas, Anallely, Méndez-Pérez, Alejandra, Méndez-Galván, Jorge, Astudillo-de la Vega, Horacio, Gabiño-López, Belem, Nava-Monroy, Ricardo, López-Flores a la Torre, Alejandro, López-Vergara Anaya, Tanit, Vilar-Compte, Diana, Coquis-Navarrete, Uriel, Valdés-Reyes, Melissa, Sánchez-Montes, Sokani, Becker, Ingeborg
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8372963/
https://www.ncbi.nlm.nih.gov/pubmed/34407100
http://dx.doi.org/10.1371/journal.pone.0255807
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author Miranda-Ortiz, Haydee
Fernández-Figueroa, Edith A.
Ruíz-García, Erika B.
Muñoz-Rivas, Anallely
Méndez-Pérez, Alejandra
Méndez-Galván, Jorge
Astudillo-de la Vega, Horacio
Gabiño-López, Belem
Nava-Monroy, Ricardo
López-Flores a la Torre, Alejandro
López-Vergara Anaya, Tanit
Vilar-Compte, Diana
Coquis-Navarrete, Uriel
Valdés-Reyes, Melissa
Sánchez-Montes, Sokani
Becker, Ingeborg
author_facet Miranda-Ortiz, Haydee
Fernández-Figueroa, Edith A.
Ruíz-García, Erika B.
Muñoz-Rivas, Anallely
Méndez-Pérez, Alejandra
Méndez-Galván, Jorge
Astudillo-de la Vega, Horacio
Gabiño-López, Belem
Nava-Monroy, Ricardo
López-Flores a la Torre, Alejandro
López-Vergara Anaya, Tanit
Vilar-Compte, Diana
Coquis-Navarrete, Uriel
Valdés-Reyes, Melissa
Sánchez-Montes, Sokani
Becker, Ingeborg
author_sort Miranda-Ortiz, Haydee
collection PubMed
description The use of saliva for the diagnosis of SARS-CoV-2 has shown to be a good alternative to nasopharyngeal swabs (NPS), since it permits self-collection, avoids the exposure of healthy persons to infected patients, reduces waiting times, eliminates the need of personal protective equipment and is non-invasive. Yet current saliva testing is still expensive due to the need of specialized tubes containing buffers to stabilize the RNA of SARS-CoV-2 and inactivate the virus. These tubes are expensive and not always accessible in sufficient quantities. We now developed an alternative saliva testing method, using TRIzol for extraction, viral inactivation, and storage of SARS-CoV-2 RNA, combined with RT-qPCR, which was comparable in its performance to NPS. Paired saliva samples and NPS were taken from 15 asymptomatic healthcare workers and one patient with SARS-CoV-2. Further 13 patients with SARS-CoV-2 were only saliva-tested. All the tests were performed according to CDC 2019-Novel Coronavirus (2019-nCoV) Real-Time RT-PCR Diagnostic Panel. Saliva (4 mL) was taken in sterile 50 mL tubes, 1.5 mL TRIzol were added and mixed. Our results show that 5 μL of saliva RNA extracted with TRIzol allow for an adequate detection of the virus in patients positive for SARS-CoV-2 and was equally sensitive to NPS in TRIzol. We conclude that saliva testing using TRIzol is a recommendable method for diagnosis of SARS-CoV-2 since it has several advantages over currently used saliva tests: it can be done with normal sterile tubes, does not need cold-chain handling, is stable at room temperature, is non-invasive and less costly, making it more accessible for low-income countries. Cheaper saliva testing using TRIzol is especially relevant for low-income countries to optimize diagnosis and help define quarantine durations for families, healthcare workers, schools, and other public workplaces, thus decreasing infections and mortality caused by SARS-CoV-2.
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spelling pubmed-83729632021-08-19 Development of an alternative saliva test for diagnosis of SARS-CoV-2 using TRIzol: Adapting to countries with lower incomes looking for a large-scale detection program Miranda-Ortiz, Haydee Fernández-Figueroa, Edith A. Ruíz-García, Erika B. Muñoz-Rivas, Anallely Méndez-Pérez, Alejandra Méndez-Galván, Jorge Astudillo-de la Vega, Horacio Gabiño-López, Belem Nava-Monroy, Ricardo López-Flores a la Torre, Alejandro López-Vergara Anaya, Tanit Vilar-Compte, Diana Coquis-Navarrete, Uriel Valdés-Reyes, Melissa Sánchez-Montes, Sokani Becker, Ingeborg PLoS One Research Article The use of saliva for the diagnosis of SARS-CoV-2 has shown to be a good alternative to nasopharyngeal swabs (NPS), since it permits self-collection, avoids the exposure of healthy persons to infected patients, reduces waiting times, eliminates the need of personal protective equipment and is non-invasive. Yet current saliva testing is still expensive due to the need of specialized tubes containing buffers to stabilize the RNA of SARS-CoV-2 and inactivate the virus. These tubes are expensive and not always accessible in sufficient quantities. We now developed an alternative saliva testing method, using TRIzol for extraction, viral inactivation, and storage of SARS-CoV-2 RNA, combined with RT-qPCR, which was comparable in its performance to NPS. Paired saliva samples and NPS were taken from 15 asymptomatic healthcare workers and one patient with SARS-CoV-2. Further 13 patients with SARS-CoV-2 were only saliva-tested. All the tests were performed according to CDC 2019-Novel Coronavirus (2019-nCoV) Real-Time RT-PCR Diagnostic Panel. Saliva (4 mL) was taken in sterile 50 mL tubes, 1.5 mL TRIzol were added and mixed. Our results show that 5 μL of saliva RNA extracted with TRIzol allow for an adequate detection of the virus in patients positive for SARS-CoV-2 and was equally sensitive to NPS in TRIzol. We conclude that saliva testing using TRIzol is a recommendable method for diagnosis of SARS-CoV-2 since it has several advantages over currently used saliva tests: it can be done with normal sterile tubes, does not need cold-chain handling, is stable at room temperature, is non-invasive and less costly, making it more accessible for low-income countries. Cheaper saliva testing using TRIzol is especially relevant for low-income countries to optimize diagnosis and help define quarantine durations for families, healthcare workers, schools, and other public workplaces, thus decreasing infections and mortality caused by SARS-CoV-2. Public Library of Science 2021-08-18 /pmc/articles/PMC8372963/ /pubmed/34407100 http://dx.doi.org/10.1371/journal.pone.0255807 Text en © 2021 Miranda-Ortiz et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Miranda-Ortiz, Haydee
Fernández-Figueroa, Edith A.
Ruíz-García, Erika B.
Muñoz-Rivas, Anallely
Méndez-Pérez, Alejandra
Méndez-Galván, Jorge
Astudillo-de la Vega, Horacio
Gabiño-López, Belem
Nava-Monroy, Ricardo
López-Flores a la Torre, Alejandro
López-Vergara Anaya, Tanit
Vilar-Compte, Diana
Coquis-Navarrete, Uriel
Valdés-Reyes, Melissa
Sánchez-Montes, Sokani
Becker, Ingeborg
Development of an alternative saliva test for diagnosis of SARS-CoV-2 using TRIzol: Adapting to countries with lower incomes looking for a large-scale detection program
title Development of an alternative saliva test for diagnosis of SARS-CoV-2 using TRIzol: Adapting to countries with lower incomes looking for a large-scale detection program
title_full Development of an alternative saliva test for diagnosis of SARS-CoV-2 using TRIzol: Adapting to countries with lower incomes looking for a large-scale detection program
title_fullStr Development of an alternative saliva test for diagnosis of SARS-CoV-2 using TRIzol: Adapting to countries with lower incomes looking for a large-scale detection program
title_full_unstemmed Development of an alternative saliva test for diagnosis of SARS-CoV-2 using TRIzol: Adapting to countries with lower incomes looking for a large-scale detection program
title_short Development of an alternative saliva test for diagnosis of SARS-CoV-2 using TRIzol: Adapting to countries with lower incomes looking for a large-scale detection program
title_sort development of an alternative saliva test for diagnosis of sars-cov-2 using trizol: adapting to countries with lower incomes looking for a large-scale detection program
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8372963/
https://www.ncbi.nlm.nih.gov/pubmed/34407100
http://dx.doi.org/10.1371/journal.pone.0255807
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