Evaluation of SYBR Green real time PCR for detecting SARS-CoV-2 from clinical samples

The pandemic caused by SARS-CoV-2 has triggered an extraordinary collapse of healthcare systems and hundred thousand of deaths worldwide. Following the declaration of the outbreak as a Public Health Emergency of International Concern by the World Health Organization (WHO) on January 30th, 2020, it h...

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Autores: Pereira Gómez, Marianoel, Fajardo Rossi, Álvaro, Echeverría Chagas, Natalia, López Tort, Luis Fernando, Perbolianachis Duarte, Paula, Costábile Cristech, Alicia, Aldunate Caramori, Fabián, Moreno Karlen, María del Pilar, Moratorio, Gonzalo
Tipo de documento: artigo
Estado:Versão publicada
Data de publicação:2021
País:Uruguay
Recursos:Universidad de la República
Repositório:COLIBRI
Idioma:inglês
OAI Identifier:oai:colibri.udelar.edu.uy:20.500.12008/33221
Acesso em linha:https://hdl.handle.net/20.500.12008/33221
Access Level:Acceso aberto
Palavra-chave:SARS-CoV-2
Molecular detection
SYBR Green
RT-qPCR
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dc.title.none.fl_str_mv Evaluation of SYBR Green real time PCR for detecting SARS-CoV-2 from clinical samples
title Evaluation of SYBR Green real time PCR for detecting SARS-CoV-2 from clinical samples
spellingShingle Evaluation of SYBR Green real time PCR for detecting SARS-CoV-2 from clinical samples
Pereira Gómez, Marianoel
SARS-CoV-2
Molecular detection
SYBR Green
RT-qPCR
title_short Evaluation of SYBR Green real time PCR for detecting SARS-CoV-2 from clinical samples
title_full Evaluation of SYBR Green real time PCR for detecting SARS-CoV-2 from clinical samples
title_fullStr Evaluation of SYBR Green real time PCR for detecting SARS-CoV-2 from clinical samples
title_full_unstemmed Evaluation of SYBR Green real time PCR for detecting SARS-CoV-2 from clinical samples
title_sort Evaluation of SYBR Green real time PCR for detecting SARS-CoV-2 from clinical samples
dc.creator.none.fl_str_mv Pereira Gómez, Marianoel
Fajardo Rossi, Álvaro
Echeverría Chagas, Natalia
López Tort, Luis Fernando
Perbolianachis Duarte, Paula
Costábile Cristech, Alicia
Aldunate Caramori, Fabián
Moreno Karlen, María del Pilar
Moratorio, Gonzalo
author Pereira Gómez, Marianoel
author_facet Pereira Gómez, Marianoel
Fajardo Rossi, Álvaro
Echeverría Chagas, Natalia
López Tort, Luis Fernando
Perbolianachis Duarte, Paula
Costábile Cristech, Alicia
Aldunate Caramori, Fabián
Moreno Karlen, María del Pilar
Moratorio, Gonzalo
author_role author
author2 Fajardo Rossi, Álvaro
Echeverría Chagas, Natalia
López Tort, Luis Fernando
Perbolianachis Duarte, Paula
Costábile Cristech, Alicia
Aldunate Caramori, Fabián
Moreno Karlen, María del Pilar
Moratorio, Gonzalo
author2_role author
author
author
author
author
author
author
author
dc.contributor.filiacion.none.fl_str_mv Pereira Gómez Marianoel, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares.
Fajardo Rossi Álvaro, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares.
Echeverría Chagas Natalia, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares.
López Tort Luis Fernando, Universidad de la República (Uruguay). CENUR
Perbolianachis Duarte Paula, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares.
Costábile Cristech Alicia, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares.
Aldunate Caramori Fabián, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares.
Moreno Karlen María del Pilar, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares.
Moratorio Gonzalo, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares.
dc.subject.es.fl_str_mv SARS-CoV-2
Molecular detection
SYBR Green
RT-qPCR
topic SARS-CoV-2
Molecular detection
SYBR Green
RT-qPCR
description The pandemic caused by SARS-CoV-2 has triggered an extraordinary collapse of healthcare systems and hundred thousand of deaths worldwide. Following the declaration of the outbreak as a Public Health Emergency of International Concern by the World Health Organization (WHO) on January 30th, 2020, it has become imperative to develop diagnostic tools to reliably detect the virus in infected patients. Several methods based on real time reverse transcription polymerase chain reaction (RT-qPCR) for the detection of SARS-CoV-2 genomic RNA have been developed. In addition, these methods have been recommended by the WHO for laboratory diagnosis. Since most of these protocols are based on the use of fluorogenic probes and one-step reagents (cDNA synthesis followed by PCR amplification in the same tube), these techniques can be difficult to perform given the limited supply of reagents in low- and middle-income countries. In order to develop an inexpensive SARS-CoV-2 detection protocol using available resources we evaluated the SYBR Green based detection of SARS-CoV-2 to establish a suitable assay. To do so, we adapted one of the WHO recommended TaqMan-based one-step real time PCR protocols (from the University of Hong Kong) to SYBR Green. Our results indicate that SYBR-Green detection of ORF1b-nsp14 target represents a reliable cost-effective alternative to increase the testing capacity.
publishDate 2021
dc.date.issued.none.fl_str_mv 2021
dc.date.accessioned.none.fl_str_mv 2022-08-19T13:05:41Z
dc.date.available.none.fl_str_mv 2022-08-19T13:05:41Z
dc.type.es.fl_str_mv Artículo
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dc.identifier.citation.es.fl_str_mv Pereira Gómez, M, Fajardo Rossi, Á, Echeverría Chagas, N [y otros autores]. "Evaluation of SYBR Green real time PCR for detecting SARS-CoV-2 from clinical samples". Journal of Virological Methods. [en línea] 2021, 289: 114035. 8 h. Doi: 10.1016/j.jviromet.2020.114035.
dc.identifier.issn.none.fl_str_mv 0166-0934
dc.identifier.uri.none.fl_str_mv https://hdl.handle.net/20.500.12008/33221
dc.identifier.doi.none.fl_str_mv 10.1016/j.jviromet.2020.114035
identifier_str_mv Pereira Gómez, M, Fajardo Rossi, Á, Echeverría Chagas, N [y otros autores]. "Evaluation of SYBR Green real time PCR for detecting SARS-CoV-2 from clinical samples". Journal of Virological Methods. [en línea] 2021, 289: 114035. 8 h. Doi: 10.1016/j.jviromet.2020.114035.
0166-0934
10.1016/j.jviromet.2020.114035
url https://hdl.handle.net/20.500.12008/33221
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eng
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language eng
dc.relation.none.fl_str_mv Journal of Virological Methods, 2021, 289: 114035.
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eu_rights_str_mv openAccess
rights_invalid_str_mv Licencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0)
dc.format.extent.es.fl_str_mv 8 h
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dc.publisher.es.fl_str_mv Elsevier
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spelling Pereira Gómez Marianoel, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares.Fajardo Rossi Álvaro, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares.Echeverría Chagas Natalia, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares.López Tort Luis Fernando, Universidad de la República (Uruguay). CENURPerbolianachis Duarte Paula, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares.Costábile Cristech Alicia, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares.Aldunate Caramori Fabián, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares.Moreno Karlen María del Pilar, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares.Moratorio Gonzalo, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares.2022-08-19T13:05:41Z2022-08-19T13:05:41Z2021Pereira Gómez, M, Fajardo Rossi, Á, Echeverría Chagas, N [y otros autores]. "Evaluation of SYBR Green real time PCR for detecting SARS-CoV-2 from clinical samples". Journal of Virological Methods. [en línea] 2021, 289: 114035. 8 h. Doi: 10.1016/j.jviromet.2020.114035.0166-0934https://hdl.handle.net/20.500.12008/3322110.1016/j.jviromet.2020.114035The pandemic caused by SARS-CoV-2 has triggered an extraordinary collapse of healthcare systems and hundred thousand of deaths worldwide. Following the declaration of the outbreak as a Public Health Emergency of International Concern by the World Health Organization (WHO) on January 30th, 2020, it has become imperative to develop diagnostic tools to reliably detect the virus in infected patients. Several methods based on real time reverse transcription polymerase chain reaction (RT-qPCR) for the detection of SARS-CoV-2 genomic RNA have been developed. In addition, these methods have been recommended by the WHO for laboratory diagnosis. Since most of these protocols are based on the use of fluorogenic probes and one-step reagents (cDNA synthesis followed by PCR amplification in the same tube), these techniques can be difficult to perform given the limited supply of reagents in low- and middle-income countries. In order to develop an inexpensive SARS-CoV-2 detection protocol using available resources we evaluated the SYBR Green based detection of SARS-CoV-2 to establish a suitable assay. To do so, we adapted one of the WHO recommended TaqMan-based one-step real time PCR protocols (from the University of Hong Kong) to SYBR Green. Our results indicate that SYBR-Green detection of ORF1b-nsp14 target represents a reliable cost-effective alternative to increase the testing capacity.Submitted by Parodi Mónica (mparodi@fcien.edu.uy) on 2022-08-12T15:00:46Z No. of bitstreams: 2 license_rdf: 23149 bytes, checksum: 1996b8461bc290aef6a27d78c67b6b52 (MD5) 101016jjviromet2020114035.pdf: 2231401 bytes, checksum: 358ec618dbf7e0155ef57384762d6f8f (MD5)Approved for entry into archive by Faget Cecilia (lfaget@fcien.edu.uy) on 2022-08-19T11:39:23Z (GMT) No. of bitstreams: 2 license_rdf: 23149 bytes, checksum: 1996b8461bc290aef6a27d78c67b6b52 (MD5) 101016jjviromet2020114035.pdf: 2231401 bytes, checksum: 358ec618dbf7e0155ef57384762d6f8f (MD5)Made available in DSpace by Luna Fabiana (fabiana.luna@seciu.edu.uy) on 2022-08-19T13:05:41Z (GMT). No. of bitstreams: 2 license_rdf: 23149 bytes, checksum: 1996b8461bc290aef6a27d78c67b6b52 (MD5) 101016jjviromet2020114035.pdf: 2231401 bytes, checksum: 358ec618dbf7e0155ef57384762d6f8f (MD5) Previous issue date: 2021ANII: FCE_1_2019_1_156157ANII: FCE_1_2019_1_1559308 happlication/pdfenengElsevierJournal of Virological Methods, 2021, 289: 114035.Las obras depositadas en el Repositorio se rigen por la Ordenanza de los Derechos de la Propiedad Intelectual de la Universidad de la República.(Res. Nº 91 de C.D.C. de 8/III/1994 – D.O. 7/IV/1994) y por la Ordenanza del Repositorio Abierto de la Universidad de la República (Res. Nº 16 de C.D.C. de 07/10/2014)info:eu-repo/semantics/openAccessLicencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0)SARS-CoV-2Molecular detectionSYBR GreenRT-qPCREvaluation of SYBR Green real time PCR for detecting SARS-CoV-2 from clinical samplesArtículoinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionreponame:COLIBRIinstname:Universidad de la Repúblicainstacron:Universidad de la RepúblicaPereira Gómez, MarianoelFajardo Rossi, ÁlvaroEcheverría Chagas, NataliaLópez Tort, Luis FernandoPerbolianachis Duarte, PaulaCostábile Cristech, AliciaAldunate Caramori, FabiánMoreno Karlen, María del PilarMoratorio, GonzaloLICENSElicense.txtlicense.txttext/plain; charset=utf-84267http://localhost:8080/xmlui/bitstream/20.500.12008/33221/5/license.txt6429389a7df7277b72b7924fdc7d47a9MD55CC-LICENSElicense_urllicense_urltext/plain; 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públicahttps://udelar.edu.uy/https://www.colibri.udelar.edu.uy/oai/requestkarina.camps@seciu.edu.uyUruguayopendoar:47712022-08-19T13:05:41COLIBRI - Universidad de la Repúblicafalse
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