Assessment of an in-house IgG ELISA targeting SARS-CoV-2 RBD: Applications in infected and vaccinated individuals

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da Costa, Hernan Hermes Monteiro ; Silva, Valeria Oliveira ; Amorim, Gustavo Carvalho ; Guereschi, Marcia Grando ; Sergio, Luciana Marciano ; Gomes, Carlos Henrique Rodrigues ; Hong, Marisa Ailin ; de Oliveira, Elaine Lopes ; Brigido, Luis Fernando de Macedo ; Lindoso, Jose Angelo Lauletta ; Prudencio, Carlos Roberto

• Document type: Journal article
• Source: JOURNAL OF IMMUNOLOGICAL METHODS; v. 530, p. 12-pg., 2024-05-22.

Abstract

The study evoluated an in-house Spike Receptor Binding Domain Enzyme-Linked Immunosorbent Assay (RBDIgG-ELISA) for detecting SARS-CoV-2 IgG antibodies in infected and vaccinated individuals. The assay demonstrated a sensitivity of 91%, specificity of 99.25%, and accuracy of 95.13%. Precision and reproducibility were highly consistent. The RBD-IgG-ELISA was able to detect 96.25% of Polymerase chain reaction (PCR) confirmed cases for SARS-CoV-2 infection, demonstrating positive and negative predictive values of 99,18% and 91,69%, respectively. In an epidemiological survey, ELISA, lateral flow immunochromatographic assay (LFIA), and electrochemiluminescence immunoassay (ECLIA) exhibited diagnostic sensitivities of 68.29%, 63.41%, and 70.73%, respectively, along with specificities of 82.93%, 80.49%, and 80.49%, respectively. Agreement between RBD-IgG-ELISA/PCR was moderate (k index 0.512). However, good agreement between different assays (RBDIgG-ELISA/LFIA k index 0.875, RBD-IgG-ELISA/ECLIA k index 0.901). Test performance on individuals' samples were inferior due to seroconversion time and chronicity. The IgG-RBD-ELISA assay demonstrated its effectiveness in monitoring antibody levels among healthcare professionals, revealing significant differences both before and after the administration of the third vaccine dose, with heightened protection levels observed following the third dose in five Coronavirus disease (COVID-19) vaccine regimens. In conclusion, the RBD-IgG-ELISA exhibits high reproducibility, specificity, and sensitivity, making it a suitable assay validated for serosurveillance and for obtaining information about COVID-19 infections or vaccinations. (AU)

• FAPESP's process: 23/13212-8 - Assessment of the humoral immune response after the bivalent mRNA vaccine against SARS-Cov-2 through the PhIP-Seq technique
• Grantee: Carlos Roberto Prudencio
• Support Opportunities: Regular Research Grants
• FAPESP's process: 21/11936-3 - Center for Translational Science and Biopharmaceutical Development
• Grantee: Benedito Barraviera
• Support Opportunities: Research Grants - Science Centers for Development
• FAPESP's process: 23/06453-9 - Construction and selection of a scFv monoclonal antibody library against the spike protein of SARS-CoV-2 variants of concern by Phage Display technology
• Grantee: Carlos Henrique Rodrigues Gomes
• Support Opportunities: Scholarships in Brazil - Technical Training Program - Technical Training
• FAPESP's process: 22/05566-1 - Construction and selection of a library of scFv monoclonal antibodies against the SARS-CoV-2 spike protein by Phage Display technology
• Grantee: Hernan Hermes Monteiro da Costa
• Support Opportunities: Scholarships in Brazil - Doctorate