High sensitivity-low cost detection of SARS-CoV-2 by two steps end point RT-PCR with agarose gel electrophoresis visualization

Abstract More than one year since Coronavirus disease 2019 (COVID-19) pandemic outbreak, the gold standard technique for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection is still the RT-qPCR. This is a limitation to increase testing capacities, particularly at developing countr...

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Autores principales:	Solange Figueroa, Byron Freire-Paspuel, Patricio Vega-Mariño, Alberto Velez, Marilyn Cruz, Washington B. Cardenas, Miguel Angel Garcia-Bereguiain
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/717de7efe5ce4400995f289132714e92
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spelling	oai:doaj.org-article:717de7efe5ce4400995f289132714e922021-11-08T10:48:03ZHigh sensitivity-low cost detection of SARS-CoV-2 by two steps end point RT-PCR with agarose gel electrophoresis visualization10.1038/s41598-021-00900-82045-2322https://doaj.org/article/717de7efe5ce4400995f289132714e922021-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-00900-8https://doaj.org/toc/2045-2322Abstract More than one year since Coronavirus disease 2019 (COVID-19) pandemic outbreak, the gold standard technique for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection is still the RT-qPCR. This is a limitation to increase testing capacities, particularly at developing countries, as expensive reagents and equipment are required. We developed a two steps end point RT-PCR reaction with SARS-CoV-2 Nucleocapsid (N) gene and Ribonuclease P (RNase P) specific primers where viral amplicons were verified by agarose gel electrophoresis. We carried out a clinical performance and analytical sensitivity evaluation for this two-steps end point RT-PCR method with 242 nasopharyngeal samples using the CDC RT-qPCR protocol as a gold standard technique. With a specificity of 95.8%, a sensitivity of 95.1%, and a limit of detection of 20 viral RNA copies/uL, this two steps end point RT-PCR assay is an affordable and reliable method for SARS-CoV-2 detection. This protocol would allow to extend COVID-19 diagnosis to basic molecular biology laboratories with a potential positive impact in surveillance programs at developing countries.Solange FigueroaByron Freire-PaspuelPatricio Vega-MariñoAlberto VelezMarilyn CruzWashington B. CardenasMiguel Angel Garcia-BereguiainNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-5 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Solange Figueroa Byron Freire-Paspuel Patricio Vega-Mariño Alberto Velez Marilyn Cruz Washington B. Cardenas Miguel Angel Garcia-Bereguiain High sensitivity-low cost detection of SARS-CoV-2 by two steps end point RT-PCR with agarose gel electrophoresis visualization
description	Abstract More than one year since Coronavirus disease 2019 (COVID-19) pandemic outbreak, the gold standard technique for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection is still the RT-qPCR. This is a limitation to increase testing capacities, particularly at developing countries, as expensive reagents and equipment are required. We developed a two steps end point RT-PCR reaction with SARS-CoV-2 Nucleocapsid (N) gene and Ribonuclease P (RNase P) specific primers where viral amplicons were verified by agarose gel electrophoresis. We carried out a clinical performance and analytical sensitivity evaluation for this two-steps end point RT-PCR method with 242 nasopharyngeal samples using the CDC RT-qPCR protocol as a gold standard technique. With a specificity of 95.8%, a sensitivity of 95.1%, and a limit of detection of 20 viral RNA copies/uL, this two steps end point RT-PCR assay is an affordable and reliable method for SARS-CoV-2 detection. This protocol would allow to extend COVID-19 diagnosis to basic molecular biology laboratories with a potential positive impact in surveillance programs at developing countries.
format	article
author	Solange Figueroa Byron Freire-Paspuel Patricio Vega-Mariño Alberto Velez Marilyn Cruz Washington B. Cardenas Miguel Angel Garcia-Bereguiain
author_facet	Solange Figueroa Byron Freire-Paspuel Patricio Vega-Mariño Alberto Velez Marilyn Cruz Washington B. Cardenas Miguel Angel Garcia-Bereguiain
author_sort	Solange Figueroa
title	High sensitivity-low cost detection of SARS-CoV-2 by two steps end point RT-PCR with agarose gel electrophoresis visualization
title_short	High sensitivity-low cost detection of SARS-CoV-2 by two steps end point RT-PCR with agarose gel electrophoresis visualization
title_full	High sensitivity-low cost detection of SARS-CoV-2 by two steps end point RT-PCR with agarose gel electrophoresis visualization
title_fullStr	High sensitivity-low cost detection of SARS-CoV-2 by two steps end point RT-PCR with agarose gel electrophoresis visualization
title_full_unstemmed	High sensitivity-low cost detection of SARS-CoV-2 by two steps end point RT-PCR with agarose gel electrophoresis visualization
title_sort	high sensitivity-low cost detection of sars-cov-2 by two steps end point rt-pcr with agarose gel electrophoresis visualization
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/717de7efe5ce4400995f289132714e92
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High sensitivity-low cost detection of SARS-CoV-2 by two steps end point RT-PCR with agarose gel electrophoresis visualization

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