SARS-CoV-2 Rapid Antigen Test Based on a New Anti-Nucleocapsid Protein Monoclonal Antibody: Development and Real-Time Validation
SARS-CoV-2 diagnostic tests have become an important tool for pandemic control. Among the alternatives for COVID-19 diagnosis, antigen rapid diagnostic tests (Ag-RDT) are very convenient and widely used. However, as SARS-CoV-2 variants may continuously emerge, the replacement of tests and reagents m...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-09-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/11/10/2422 |
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| author | Fabiana Fioravante Coelho Miriam Aparecida da Silva Thiciany Blener Lopes Juliana Moutinho Polatto Natália Salazar de Castro Luis Adan Flores Andrade Karine Lima Lourenço Hugo Itaru Sato Alex Fiorini de Carvalho Helena Perez Coelho Flávia Fonseca Bagno Daniela Luz Vincent Louis Viala Pedro Queiroz Cattony Bruna de Sousa Melo Ana Maria Moro Wagner Quintilio Ana Paula Barbosa Camila Gasque Bomfim Camila Pereira Soares Cristiane Rodrigues Guzzo Flavio Guimarães Fonseca Edison Luiz Durigon Ricardo Tostes Gazzinelli Santuza M. Ribeiro Teixeira Roxane Maria Fontes Piazza Ana Paula Fernandes |
| author_facet | Fabiana Fioravante Coelho Miriam Aparecida da Silva Thiciany Blener Lopes Juliana Moutinho Polatto Natália Salazar de Castro Luis Adan Flores Andrade Karine Lima Lourenço Hugo Itaru Sato Alex Fiorini de Carvalho Helena Perez Coelho Flávia Fonseca Bagno Daniela Luz Vincent Louis Viala Pedro Queiroz Cattony Bruna de Sousa Melo Ana Maria Moro Wagner Quintilio Ana Paula Barbosa Camila Gasque Bomfim Camila Pereira Soares Cristiane Rodrigues Guzzo Flavio Guimarães Fonseca Edison Luiz Durigon Ricardo Tostes Gazzinelli Santuza M. Ribeiro Teixeira Roxane Maria Fontes Piazza Ana Paula Fernandes |
| author_sort | Fabiana Fioravante Coelho |
| collection | DOAJ |
| description | SARS-CoV-2 diagnostic tests have become an important tool for pandemic control. Among the alternatives for COVID-19 diagnosis, antigen rapid diagnostic tests (Ag-RDT) are very convenient and widely used. However, as SARS-CoV-2 variants may continuously emerge, the replacement of tests and reagents may be required to maintain the sensitivity of Ag-RDTs. Here, we describe the development and validation of an Ag-RDT during an outbreak of the Omicron variant, including the characterization of a new monoclonal antibody (anti-DTC-N 1B3 mAb) that recognizes the Nucleocapsid protein (N). The anti-DTC-N 1B3 mAb recognized the sequence TFPPTEPKKDKKK located at the C-terminus of the N protein of main SARS-CoV-2 variants of concern. Accordingly, the Ag-RDT prototypes using the anti-DTC-N 1B3 mAB detected all the SARS-CoV-2 variants—Wuhan, Alpha, Gamma, Delta, P2 and Omicron. The performance of the best prototype (sensitivity of 95.2% for samples with Ct ≤ 25; specificity of 98.3% and overall accuracy of 85.0%) met the WHO recommendations. Moreover, results from a patients’ follow-up study indicated that, if performed within the first three days after onset of symptoms, the Ag-RDT displayed 100% sensitivity. Thus, the new mAb and the Ag-RDT developed herein may constitute alternative tools for COVID-19 point-of-care diagnosis and epidemiological surveillance. |
| first_indexed | 2024-03-10T21:02:47Z |
| format | Article |
| id | doaj.art-9d2b5f4abaf046d7a458a65c8a0c962b |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-10T21:02:47Z |
| publishDate | 2023-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-9d2b5f4abaf046d7a458a65c8a0c962b2023-11-19T17:26:30ZengMDPI AGMicroorganisms2076-26072023-09-011110242210.3390/microorganisms11102422SARS-CoV-2 Rapid Antigen Test Based on a New Anti-Nucleocapsid Protein Monoclonal Antibody: Development and Real-Time ValidationFabiana Fioravante Coelho0Miriam Aparecida da Silva1Thiciany Blener Lopes2Juliana Moutinho Polatto3Natália Salazar de Castro4Luis Adan Flores Andrade5Karine Lima Lourenço6Hugo Itaru Sato7Alex Fiorini de Carvalho8Helena Perez Coelho9Flávia Fonseca Bagno10Daniela Luz11Vincent Louis Viala12Pedro Queiroz Cattony13Bruna de Sousa Melo14Ana Maria Moro15Wagner Quintilio16Ana Paula Barbosa17Camila Gasque Bomfim18Camila Pereira Soares19Cristiane Rodrigues Guzzo20Flavio Guimarães Fonseca21Edison Luiz Durigon22Ricardo Tostes Gazzinelli23Santuza M. Ribeiro Teixeira24Roxane Maria Fontes Piazza25Ana Paula Fernandes26Faculdade de Farmácia, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, BrazilInstituto Butantan, São Paulo 05503-900, BrazilCentro de Tecnologia em Vacinas, Universidade Federal de Minas Gerais, Belo Horizonte 31310-260, BrazilInstituto Butantan, São Paulo 05503-900, BrazilCentro de Tecnologia em Vacinas, Universidade Federal de Minas Gerais, Belo Horizonte 31310-260, BrazilCentro de Tecnologia em Vacinas, Universidade Federal de Minas Gerais, Belo Horizonte 31310-260, BrazilCentro de Tecnologia em Vacinas, Universidade Federal de Minas Gerais, Belo Horizonte 31310-260, BrazilCentro de Tecnologia em Vacinas, Universidade Federal de Minas Gerais, Belo Horizonte 31310-260, BrazilCentro de Tecnologia em Vacinas, Universidade Federal de Minas Gerais, Belo Horizonte 31310-260, BrazilCentro de Tecnologia em Vacinas, Universidade Federal de Minas Gerais, Belo Horizonte 31310-260, BrazilCentro de Tecnologia em Vacinas, Universidade Federal de Minas Gerais, Belo Horizonte 31310-260, BrazilInstituto Butantan, São Paulo 05503-900, BrazilInstituto Butantan, São Paulo 05503-900, BrazilInstituto Butantan, São Paulo 05503-900, BrazilInstituto Butantan, São Paulo 05503-900, BrazilInstituto Butantan, São Paulo 05503-900, BrazilInstituto Butantan, São Paulo 05503-900, BrazilInstituto Butantan, São Paulo 05503-900, BrazilDepartment of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo 05508-900, BrazilDepartment of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo 05508-900, BrazilDepartment of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo 05508-900, BrazilCentro de Tecnologia em Vacinas, Universidade Federal de Minas Gerais, Belo Horizonte 31310-260, BrazilDepartment of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo 05508-900, BrazilCentro de Tecnologia em Vacinas, Universidade Federal de Minas Gerais, Belo Horizonte 31310-260, BrazilCentro de Tecnologia em Vacinas, Universidade Federal de Minas Gerais, Belo Horizonte 31310-260, BrazilInstituto Butantan, São Paulo 05503-900, BrazilFaculdade de Farmácia, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, BrazilSARS-CoV-2 diagnostic tests have become an important tool for pandemic control. Among the alternatives for COVID-19 diagnosis, antigen rapid diagnostic tests (Ag-RDT) are very convenient and widely used. However, as SARS-CoV-2 variants may continuously emerge, the replacement of tests and reagents may be required to maintain the sensitivity of Ag-RDTs. Here, we describe the development and validation of an Ag-RDT during an outbreak of the Omicron variant, including the characterization of a new monoclonal antibody (anti-DTC-N 1B3 mAb) that recognizes the Nucleocapsid protein (N). The anti-DTC-N 1B3 mAb recognized the sequence TFPPTEPKKDKKK located at the C-terminus of the N protein of main SARS-CoV-2 variants of concern. Accordingly, the Ag-RDT prototypes using the anti-DTC-N 1B3 mAB detected all the SARS-CoV-2 variants—Wuhan, Alpha, Gamma, Delta, P2 and Omicron. The performance of the best prototype (sensitivity of 95.2% for samples with Ct ≤ 25; specificity of 98.3% and overall accuracy of 85.0%) met the WHO recommendations. Moreover, results from a patients’ follow-up study indicated that, if performed within the first three days after onset of symptoms, the Ag-RDT displayed 100% sensitivity. Thus, the new mAb and the Ag-RDT developed herein may constitute alternative tools for COVID-19 point-of-care diagnosis and epidemiological surveillance.https://www.mdpi.com/2076-2607/11/10/2422SARS-CoV-2diagnosisnucleocapsid (N) antigenIgG2b monoclonal antibodyAg-RDT developmentvalidation |
| spellingShingle | Fabiana Fioravante Coelho Miriam Aparecida da Silva Thiciany Blener Lopes Juliana Moutinho Polatto Natália Salazar de Castro Luis Adan Flores Andrade Karine Lima Lourenço Hugo Itaru Sato Alex Fiorini de Carvalho Helena Perez Coelho Flávia Fonseca Bagno Daniela Luz Vincent Louis Viala Pedro Queiroz Cattony Bruna de Sousa Melo Ana Maria Moro Wagner Quintilio Ana Paula Barbosa Camila Gasque Bomfim Camila Pereira Soares Cristiane Rodrigues Guzzo Flavio Guimarães Fonseca Edison Luiz Durigon Ricardo Tostes Gazzinelli Santuza M. Ribeiro Teixeira Roxane Maria Fontes Piazza Ana Paula Fernandes SARS-CoV-2 Rapid Antigen Test Based on a New Anti-Nucleocapsid Protein Monoclonal Antibody: Development and Real-Time Validation Microorganisms SARS-CoV-2 diagnosis nucleocapsid (N) antigen IgG2b monoclonal antibody Ag-RDT development validation |
| title | SARS-CoV-2 Rapid Antigen Test Based on a New Anti-Nucleocapsid Protein Monoclonal Antibody: Development and Real-Time Validation |
| title_full | SARS-CoV-2 Rapid Antigen Test Based on a New Anti-Nucleocapsid Protein Monoclonal Antibody: Development and Real-Time Validation |
| title_fullStr | SARS-CoV-2 Rapid Antigen Test Based on a New Anti-Nucleocapsid Protein Monoclonal Antibody: Development and Real-Time Validation |
| title_full_unstemmed | SARS-CoV-2 Rapid Antigen Test Based on a New Anti-Nucleocapsid Protein Monoclonal Antibody: Development and Real-Time Validation |
| title_short | SARS-CoV-2 Rapid Antigen Test Based on a New Anti-Nucleocapsid Protein Monoclonal Antibody: Development and Real-Time Validation |
| title_sort | sars cov 2 rapid antigen test based on a new anti nucleocapsid protein monoclonal antibody development and real time validation |
| topic | SARS-CoV-2 diagnosis nucleocapsid (N) antigen IgG2b monoclonal antibody Ag-RDT development validation |
| url | https://www.mdpi.com/2076-2607/11/10/2422 |
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