Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS‑CoV‑2 seroconversion
| Main Author: | |
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| Publication Date: | 2021 |
| Other Authors: | , , , , , , , , , , , , , , , , , , |
| Format: | Article |
| Language: | por |
| Source: | Repositório Institucional da FIOCRUZ (ARCA) |
| Download full: | https://www.arca.fiocruz.br/handle/icict/49817 |
Summary: | Universidade Federal do Paraná. Setor Litoral. Matinhos, PR, Brasil. |
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Conzentino, Marcelo S.Forchhammer, KarlSouza, Emanuel Maltempi dePedrosa, Fábio O.Nogueira, Meri B.Raboni, Sônia M.Rego, Fabiane G. M.Zanette, Dalila LucíolaAoki, Mateus NóbregaNardin, Jeanine M.Fornazari, BrunaMorales, Hugo M. P.Celedon, Paola Alejandra FioraniLima, Carla V. P.Mattar, Sibelle B.Lin, Vanessa H.Morello, Luis GustavoMarchini, Fabrício KleryntonReis, Rodrigo A.Huergo, Luciano F.2021-11-12T15:24:25Z2021-11-12T15:24:25Z2021CONZENTINO, Marcelo S. et al. Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS‑CoV‑2 seroconversion. Brazilian Journal of Microbiology. p. 1–5, 2021.1678-4405https://www.arca.fiocruz.br/handle/icict/4981710.1007/s42770-021-00556-6porSociedade Brasileira de MicrobiologiaSARS-CoV-2COVID-19ELISAProteínas do NucleocapsídeoImmunologic TestsEnzyme-Linked Immunosorbent AssayNucleocapsid ProteinsPruebas InmunológicasEnsayo de Inmunoadsorción EnzimáticaProteínas de la NucleocápsideTests immunologiquesTest ELISAProtéines nucléocapsideTestes ImunológicosEnsaio de Imunoadsorção EnzimáticaAntigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS‑CoV‑2 seroconversioninfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleUniversidade Federal do Paraná. Setor Litoral. Matinhos, PR, Brasil.Interfakultäres Institut für Mikrobiologie und Infektionsmedizin der Eberhard-Karls. Universität Tübingen. Tübingen, Germany.Universidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brasil.Universidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brasil.Universidade Federal do Paraná. Hospital das Clínicas. Curitiba, PR, Brasil.Universidade Federal do Paraná. Hospital das Clínicas. Curitiba, PR, Brasil.Universidade Federal do Paraná. Programa de Pós-Graduação em Ciências Farmacêuticas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Hospital Erasto Gaertner. Curitiba, PR, Brasil.Hospital Erasto Gaertner. Curitiba, PR, Brasil.Hospital Erasto Gaertner. Curitiba, PR, Brasil.Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil. / Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil. / Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.Universidade Federal do Paraná. Setor Litoral. Matinhos, PR, Brasil.Universidade Federal do Paraná. Setor Litoral. Matinhos, PR, Brasil.Serological assays are important tools to identify previous exposure to SARS-CoV-2, helping to track COVID-19 cases and determine the level of humoral response to SARS-CoV-2 infections and/or immunization to future vaccines. Here, the SARS-CoV-2 nucleocapsid protein was expressed in Escherichia coli and purifed to homogeneity and high yield using a single chromatography step. The purifed SARS-CoV-2 nucleocapsid protein was used to develop an indirect enzyme-linked immunosorbent assay for the identifcation of human SARS-CoV-2 seroconverts. The assay sensitivity and specifcity were determined analyzing sera from 140 RT-qPCR-confrmed COVID-19 cases and 210 pre-pandemic controls. The assay operated with 90% sensitivity and 98% specifcity; identical accuracies were obtained in head-to-head comparison with a commercial ELISA kit. Antigen-coated plates were stable for up to 3 months at 4 °C. The ELISA method described is ready for mass production and will be an additional tool to track COVID-19 cases.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-83084https://www.arca.fiocruz.br/bitstream/icict/49817/1/license.txt783568c2893d2e25a99990b126be1772MD51ORIGINAL42770_2021_Article_ok.pdf42770_2021_Article_ok.pdfapplication/pdf1504795https://www.arca.fiocruz.br/bitstream/icict/49817/2/42770_2021_Article_ok.pdf460c54d460a720625116ce13b176fca7MD52icict/498172021-12-23 16:31:19.229oai:www.arca.fiocruz.br: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ório InstitucionalPUBhttps://www.arca.fiocruz.br/oai/requestrepositorio.arca@fiocruz.bropendoar:21352021-12-23T19:31:19Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)false |
| dc.title.pt_BR.fl_str_mv |
Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS‑CoV‑2 seroconversion |
| title |
Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS‑CoV‑2 seroconversion |
| spellingShingle |
Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS‑CoV‑2 seroconversion Conzentino, Marcelo S. SARS-CoV-2 COVID-19 ELISA Proteínas do Nucleocapsídeo Immunologic Tests Enzyme-Linked Immunosorbent Assay Nucleocapsid Proteins Pruebas Inmunológicas Ensayo de Inmunoadsorción Enzimática Proteínas de la Nucleocápside Tests immunologiques Test ELISA Protéines nucléocapside Testes Imunológicos Ensaio de Imunoadsorção Enzimática |
| title_short |
Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS‑CoV‑2 seroconversion |
| title_full |
Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS‑CoV‑2 seroconversion |
| title_fullStr |
Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS‑CoV‑2 seroconversion |
| title_full_unstemmed |
Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS‑CoV‑2 seroconversion |
| title_sort |
Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS‑CoV‑2 seroconversion |
| author |
Conzentino, Marcelo S. |
| author_facet |
Conzentino, Marcelo S. Forchhammer, Karl Souza, Emanuel Maltempi de Pedrosa, Fábio O. Nogueira, Meri B. Raboni, Sônia M. Rego, Fabiane G. M. Zanette, Dalila Lucíola Aoki, Mateus Nóbrega Nardin, Jeanine M. Fornazari, Bruna Morales, Hugo M. P. Celedon, Paola Alejandra Fiorani Lima, Carla V. P. Mattar, Sibelle B. Lin, Vanessa H. Morello, Luis Gustavo Marchini, Fabrício Klerynton Reis, Rodrigo A. Huergo, Luciano F. |
| author_role |
author |
| author2 |
Forchhammer, Karl Souza, Emanuel Maltempi de Pedrosa, Fábio O. Nogueira, Meri B. Raboni, Sônia M. Rego, Fabiane G. M. Zanette, Dalila Lucíola Aoki, Mateus Nóbrega Nardin, Jeanine M. Fornazari, Bruna Morales, Hugo M. P. Celedon, Paola Alejandra Fiorani Lima, Carla V. P. Mattar, Sibelle B. Lin, Vanessa H. Morello, Luis Gustavo Marchini, Fabrício Klerynton Reis, Rodrigo A. Huergo, Luciano F. |
| author2_role |
author author author author author author author author author author author author author author author author author author author |
| dc.contributor.author.fl_str_mv |
Conzentino, Marcelo S. Forchhammer, Karl Souza, Emanuel Maltempi de Pedrosa, Fábio O. Nogueira, Meri B. Raboni, Sônia M. Rego, Fabiane G. M. Zanette, Dalila Lucíola Aoki, Mateus Nóbrega Nardin, Jeanine M. Fornazari, Bruna Morales, Hugo M. P. Celedon, Paola Alejandra Fiorani Lima, Carla V. P. Mattar, Sibelle B. Lin, Vanessa H. Morello, Luis Gustavo Marchini, Fabrício Klerynton Reis, Rodrigo A. Huergo, Luciano F. |
| dc.subject.other.pt_BR.fl_str_mv |
SARS-CoV-2 COVID-19 ELISA Proteínas do Nucleocapsídeo |
| topic |
SARS-CoV-2 COVID-19 ELISA Proteínas do Nucleocapsídeo Immunologic Tests Enzyme-Linked Immunosorbent Assay Nucleocapsid Proteins Pruebas Inmunológicas Ensayo de Inmunoadsorción Enzimática Proteínas de la Nucleocápside Tests immunologiques Test ELISA Protéines nucléocapside Testes Imunológicos Ensaio de Imunoadsorção Enzimática |
| dc.subject.en.pt_BR.fl_str_mv |
Immunologic Tests Enzyme-Linked Immunosorbent Assay Nucleocapsid Proteins |
| dc.subject.es.pt_BR.fl_str_mv |
Pruebas Inmunológicas Ensayo de Inmunoadsorción Enzimática Proteínas de la Nucleocápside |
| dc.subject.fr.pt_BR.fl_str_mv |
Tests immunologiques Test ELISA Protéines nucléocapside |
| dc.subject.decs.pt_BR.fl_str_mv |
Testes Imunológicos Ensaio de Imunoadsorção Enzimática |
| description |
Universidade Federal do Paraná. Setor Litoral. Matinhos, PR, Brasil. |
| publishDate |
2021 |
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2021-11-12T15:24:25Z |
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2021-11-12T15:24:25Z |
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2021 |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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| dc.identifier.citation.fl_str_mv |
CONZENTINO, Marcelo S. et al. Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS‑CoV‑2 seroconversion. Brazilian Journal of Microbiology. p. 1–5, 2021. |
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https://www.arca.fiocruz.br/handle/icict/49817 |
| dc.identifier.issn.pt_BR.fl_str_mv |
1678-4405 |
| dc.identifier.doi.none.fl_str_mv |
10.1007/s42770-021-00556-6 |
| identifier_str_mv |
CONZENTINO, Marcelo S. et al. Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS‑CoV‑2 seroconversion. Brazilian Journal of Microbiology. p. 1–5, 2021. 1678-4405 10.1007/s42770-021-00556-6 |
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https://www.arca.fiocruz.br/handle/icict/49817 |
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por |
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openAccess |
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Sociedade Brasileira de Microbiologia |
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Sociedade Brasileira de Microbiologia |
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