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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| Format: | article |
| Language: | English |
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2021
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| Online Access: | https://hdl.handle.net/20.500.12008/33221 |
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| _version_ | 1868890253307150336 |
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| author | Pereira Gómez, Marianoel |
| 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 |
| author_browse | Aldunate Caramori, Fabián Costábile Cristech, Alicia Echeverría Chagas, Natalia Fajardo Rossi, Álvaro López Tort, Luis Fernando Moratorio, Gonzalo Moreno Karlen, María del Pilar Perbolianachis Duarte, Paula 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 |
| collection | COLIBRI |
| dc.contributor.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.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 |
| dc.date.none.fl_str_mv | 2021 2022-08-19T13:05:41Z 2022-08-19T13:05:41Z |
| dc.format.none.fl_str_mv | 8 h application/pdf |
| dc.identifier.none.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. 0166-0934 https://hdl.handle.net/20.500.12008/33221 10.1016/j.jviromet.2020.114035 |
| dc.language.none.fl_str_mv | en eng |
| dc.publisher.none.fl_str_mv | Elsevier |
| dc.relation.none.fl_str_mv | Journal of Virological Methods, 2021, 289: 114035. |
| dc.rights.none.fl_str_mv | info:eu-repo/semantics/openAccess Licencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0) |
| dc.source.none.fl_str_mv | reponame:COLIBRI instname:Universidad de la República instacron:Universidad de la República |
| dc.subject.none.fl_str_mv | SARS-CoV-2 Molecular detection SYBR Green RT-qPCR |
| dc.title.none.fl_str_mv | Evaluation of SYBR Green real time PCR for detecting SARS-CoV-2 from clinical samples |
| dc.type.none.fl_str_mv | Artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
| 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. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | anni_ffc16be4e7308cf505366de584e7a4bc |
| 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 |
| instacron_str | Universidad de la República |
| institution | Universidad de la República |
| instname_str | Universidad de la República |
| language | eng |
| language_invalid_str_mv | en |
| network_acronym_str | anni |
| network_name_str | oai-lr-anni |
| oai_identifier_str | oai:colibri.udelar.edu.uy:20.500.12008/33221 |
| publishDate | 2021 |
| publishDateSort | 2021 |
| publisher.none.fl_str_mv | Elsevier |
| reponame_str | COLIBRI |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | Licencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0) |
| spelling | Evaluation of SYBR Green real time PCR for detecting SARS-CoV-2 from clinical samplesPereira Gómez, MarianoelFajardo Rossi, ÁlvaroEcheverría Chagas, NataliaLópez Tort, Luis FernandoPerbolianachis Duarte, PaulaCostábile Cristech, AliciaAldunate Caramori, FabiánMoreno Karlen, María del PilarMoratorio, GonzaloSARS-CoV-2Molecular detectionSYBR GreenRT-qPCRThe 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.ANII: FCE_1_2019_1_156157ANII: FCE_1_2019_1_155930ElsevierPereira 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.2022-08-19T13:05:41Z2022-08-19T13:05:41Z2021Artículoinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion8 happlication/pdfPereira 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.2020.114035reponame:COLIBRIinstname:Universidad de la Repúblicainstacron:Universidad de la RepúblicaenengJournal 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)oai:colibri.udelar.edu.uy:20.500.12008/332212026-04-14T10:09:55Z |
| 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 |
| status_str | publishedVersion |
| title | 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_short | 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 |
| topic | SARS-CoV-2 Molecular detection SYBR Green RT-qPCR |
| url | https://hdl.handle.net/20.500.12008/33221 |