A fast and cheap in-house magnetic bead RNA extraction method for COVID-19 diagnosis
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1016/j.jviromet.2021.114414 http://hdl.handle.net/11449/230052 |
Resumo: | COVID-19 has posed a worldwide public health challenge affecting millions of people in different countries. Rapid and efficient detection of SARS-CoV-2 is essential for pandemic control. Reverse Transcription quantitative PCR (RT-qPCR) of nasopharyngeal swabs is the gold standard method for the virus detection, but the high demand for tests has substantially increased the costs and reduced the availability of reagents, including genetic material purification kits. Thus, the present study aimed to compare two bead-based RNA extraction methods (an in-house and a commercial kit) from nasopharyngeal swabs and RT-qPCR detection of SARS-CoV-2. Twenty-five positive and five negative nasopharyngeal swab samples were subjected to extraction of nucleic acids using both methods in an automated platform. Both protocols revealed a high correlation between Cycle Quantifications (Cqs) (r = 0.99, p < 0.0001). In addition, the in-house kit was 89.5 % cheaper when compared to the mean cost of commercial RNA extraction kits. The results show that the in-house protocol is an affordable and reliable option for RNA extraction for SARS-CoV-2 detection from nasopharyngeal swabs. |
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spelling |
A fast and cheap in-house magnetic bead RNA extraction method for COVID-19 diagnosisCOVID-19 testRNA extractionRT-qPCRSARS-CoV-2COVID-19 has posed a worldwide public health challenge affecting millions of people in different countries. Rapid and efficient detection of SARS-CoV-2 is essential for pandemic control. Reverse Transcription quantitative PCR (RT-qPCR) of nasopharyngeal swabs is the gold standard method for the virus detection, but the high demand for tests has substantially increased the costs and reduced the availability of reagents, including genetic material purification kits. Thus, the present study aimed to compare two bead-based RNA extraction methods (an in-house and a commercial kit) from nasopharyngeal swabs and RT-qPCR detection of SARS-CoV-2. Twenty-five positive and five negative nasopharyngeal swab samples were subjected to extraction of nucleic acids using both methods in an automated platform. Both protocols revealed a high correlation between Cycle Quantifications (Cqs) (r = 0.99, p < 0.0001). In addition, the in-house kit was 89.5 % cheaper when compared to the mean cost of commercial RNA extraction kits. The results show that the in-house protocol is an affordable and reliable option for RNA extraction for SARS-CoV-2 detection from nasopharyngeal swabs.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)São Paulo State University (UNESP) Institute of Biotechnology (IBTEC)São Paulo State University (UNESP) Department of Animal Production and Preventive Veterinary Medicine School of Veterinary Medicine and Animal ScienceSão Paulo State University (UNESP) Department of Veterinary Clinics School of Veterinary Medicine and Animal ScienceSão Paulo State University (UNESP) Institute of Biotechnology (IBTEC)São Paulo State University (UNESP) Department of Animal Production and Preventive Veterinary Medicine School of Veterinary Medicine and Animal ScienceSão Paulo State University (UNESP) Department of Veterinary Clinics School of Veterinary Medicine and Animal ScienceFAPESP: 19/18581-6Universidade Estadual Paulista (UNESP)Possebon, Fabio Sossai [UNESP]Ullmann, Leila Sabrina [UNESP]Malossi, Camila Dantas [UNESP]Miodutzki, Gabrielle Thaís [UNESP]da Silva, Evelyn Cristine [UNESP]Machado, Eduardo Ferreira [UNESP]Braga, Iolanda Simões [UNESP]Pelaquim, Isadora Fernanda [UNESP]Araujo, João Pessoa [UNESP]2022-04-29T08:37:21Z2022-04-29T08:37:21Z2022-02-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://dx.doi.org/10.1016/j.jviromet.2021.114414Journal of Virological Methods, v. 300.1879-09840166-0934http://hdl.handle.net/11449/23005210.1016/j.jviromet.2021.1144142-s2.0-85121110711Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengJournal of Virological Methodsinfo:eu-repo/semantics/openAccess2024-09-09T13:02:02Zoai:repositorio.unesp.br:11449/230052Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestrepositoriounesp@unesp.bropendoar:29462024-09-09T13:02:02Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
A fast and cheap in-house magnetic bead RNA extraction method for COVID-19 diagnosis |
title |
A fast and cheap in-house magnetic bead RNA extraction method for COVID-19 diagnosis |
spellingShingle |
A fast and cheap in-house magnetic bead RNA extraction method for COVID-19 diagnosis Possebon, Fabio Sossai [UNESP] COVID-19 test RNA extraction RT-qPCR SARS-CoV-2 |
title_short |
A fast and cheap in-house magnetic bead RNA extraction method for COVID-19 diagnosis |
title_full |
A fast and cheap in-house magnetic bead RNA extraction method for COVID-19 diagnosis |
title_fullStr |
A fast and cheap in-house magnetic bead RNA extraction method for COVID-19 diagnosis |
title_full_unstemmed |
A fast and cheap in-house magnetic bead RNA extraction method for COVID-19 diagnosis |
title_sort |
A fast and cheap in-house magnetic bead RNA extraction method for COVID-19 diagnosis |
author |
Possebon, Fabio Sossai [UNESP] |
author_facet |
Possebon, Fabio Sossai [UNESP] Ullmann, Leila Sabrina [UNESP] Malossi, Camila Dantas [UNESP] Miodutzki, Gabrielle Thaís [UNESP] da Silva, Evelyn Cristine [UNESP] Machado, Eduardo Ferreira [UNESP] Braga, Iolanda Simões [UNESP] Pelaquim, Isadora Fernanda [UNESP] Araujo, João Pessoa [UNESP] |
author_role |
author |
author2 |
Ullmann, Leila Sabrina [UNESP] Malossi, Camila Dantas [UNESP] Miodutzki, Gabrielle Thaís [UNESP] da Silva, Evelyn Cristine [UNESP] Machado, Eduardo Ferreira [UNESP] Braga, Iolanda Simões [UNESP] Pelaquim, Isadora Fernanda [UNESP] Araujo, João Pessoa [UNESP] |
author2_role |
author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (UNESP) |
dc.contributor.author.fl_str_mv |
Possebon, Fabio Sossai [UNESP] Ullmann, Leila Sabrina [UNESP] Malossi, Camila Dantas [UNESP] Miodutzki, Gabrielle Thaís [UNESP] da Silva, Evelyn Cristine [UNESP] Machado, Eduardo Ferreira [UNESP] Braga, Iolanda Simões [UNESP] Pelaquim, Isadora Fernanda [UNESP] Araujo, João Pessoa [UNESP] |
dc.subject.por.fl_str_mv |
COVID-19 test RNA extraction RT-qPCR SARS-CoV-2 |
topic |
COVID-19 test RNA extraction RT-qPCR SARS-CoV-2 |
description |
COVID-19 has posed a worldwide public health challenge affecting millions of people in different countries. Rapid and efficient detection of SARS-CoV-2 is essential for pandemic control. Reverse Transcription quantitative PCR (RT-qPCR) of nasopharyngeal swabs is the gold standard method for the virus detection, but the high demand for tests has substantially increased the costs and reduced the availability of reagents, including genetic material purification kits. Thus, the present study aimed to compare two bead-based RNA extraction methods (an in-house and a commercial kit) from nasopharyngeal swabs and RT-qPCR detection of SARS-CoV-2. Twenty-five positive and five negative nasopharyngeal swab samples were subjected to extraction of nucleic acids using both methods in an automated platform. Both protocols revealed a high correlation between Cycle Quantifications (Cqs) (r = 0.99, p < 0.0001). In addition, the in-house kit was 89.5 % cheaper when compared to the mean cost of commercial RNA extraction kits. The results show that the in-house protocol is an affordable and reliable option for RNA extraction for SARS-CoV-2 detection from nasopharyngeal swabs. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-04-29T08:37:21Z 2022-04-29T08:37:21Z 2022-02-01 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1016/j.jviromet.2021.114414 Journal of Virological Methods, v. 300. 1879-0984 0166-0934 http://hdl.handle.net/11449/230052 10.1016/j.jviromet.2021.114414 2-s2.0-85121110711 |
url |
http://dx.doi.org/10.1016/j.jviromet.2021.114414 http://hdl.handle.net/11449/230052 |
identifier_str_mv |
Journal of Virological Methods, v. 300. 1879-0984 0166-0934 10.1016/j.jviromet.2021.114414 2-s2.0-85121110711 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Journal of Virological Methods |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
repositoriounesp@unesp.br |
_version_ |
1813546542677622784 |