Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS-CoV-2 seroconversion
| Autor: | Sonia Mara Raboni, Marcelo S Conzentino, Hugo Manuel Paz Morales, Fabiane Gomes de Moraes Rego, Bruna Fornazari, Paola Alejandra Fiorani Celedon, Carla V P Lima, Fábio O. Pedrosa, Jeanine M Nardin, Rodrigo A. Reis, Karl Forchhammer, Sibelle B Mattar, Vanessa H Lin, Luciano F. Huergo, Fabricio K Marchini, Emanuel Maltempi de Souza, Luis Gustavo Morello, Dalila Luciola Zanette, Meri Bordignon Nogueira, Mateus Nóbrega Aoki |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
medicine.medical_specialty
Coronavirus disease 2019 (COVID-19) Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viruses Immunological test Enzyme-Linked Immunosorbent Assay Biology medicine.disease_cause Antibodies Viral Microbiology Sensitivity and Specificity Serology Medical microbiology Antigen Media Technology medicine Coronavirus Nucleocapsid Proteins Humans Seroconversion Escherichia coli Nucleocapsid protein SARS-CoV-2 COVID-19 Assay sensitivity Nucleocapsid Proteins Phosphoproteins Virology Immunity Humoral ELISA Clinical Microbiology - Short Communication |
| Zdroj: | Brazilian Journal of Microbiology |
| ISSN: | 1678-4405 1517-8382 |
| Popis: | 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 purified to homogeneity and high yield using a single chromatography step. The purified SARS-CoV-2 nucleocapsid protein was used to develop an indirect enzyme-linked immunosorbent assay for the identification of human SARS-CoV-2 seroconverts. The assay sensitivity and specificity were determined analyzing sera from 140 RT-qPCR-confirmed COVID-19 cases and 210 pre-pandemic controls. The assay operated with 90% sensitivity and 98% specificity; 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. |
| Databáze: | OpenAIRE |
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