Abstract

The Covid-19 pandemic, caused by SARS-CoV-2, has resulted in over 6 million reported deaths worldwide being one of the biggest challenges the world faces today. Here we present optimizations of all steps of an enzyme-linked immunosorbent assay (ELISA)-based test to detect IgG, IgA and IgM against the trimeric spike (S) protein, receptor binding domain (RBD), and N terminal domain of the nucleocapsid (N-NTD) protein of SARS-CoV-2. We discuss how to determine specific thresholds for antibody positivity and its limitations according to the antigen used. We applied the assay to a cohort of 126 individuals from Rio de Janeiro, Brazil, consisting of 23 PCR-positive individuals and 103 individuals without a confirmed diagnosis for SARS-CoV-2 infection. To illustrate the differences in serological responses to vaccinal immunization, we applied the test in 18 individuals from our cohort before and after receiving ChAdOx-1 nCoV-19 or CoronaVac vaccines. Taken together, our results show that the test can be customized at different stages depending on its application, enabling the user to analyze different cohorts, saving time, reagents, or samples. It is also a valuable tool for elucidating the immunological consequences of new viral strains and monitoring vaccination coverage and duration of response to different immunization regimens.

Details

Title
On the caveats of a multiplex test for SARS-CoV-2 to detect seroconversion after infection or vaccination
Author
Fernandes-Siqueira, Lorena O. 1 ; Ferreira, Fabiana A. P. 1 ; Sousa, Bruna G. 1 ; Mebus-Antunes, Nathane C. 1 ; Neves-Martins, Thais C. 1 ; Almeida, Fabio C. L. 2 ; Ferreira, Gustavo C. 1 ; Salmon, Didier 1 ; Wermelinger, Luciana S. 3 ; Da Poian, Andrea T. 1 

 Federal University of Rio de Janeiro, Institute of Medical Biochemistry Leopoldo de Meis, Rio de Janeiro, Brazil (GRID:grid.8536.8) (ISNI:0000 0001 2294 473X) 
 Federal University of Rio de Janeiro, Institute of Medical Biochemistry Leopoldo de Meis, Rio de Janeiro, Brazil (GRID:grid.8536.8) (ISNI:0000 0001 2294 473X); Federal University of Rio de Janeiro, National Center for Structural Biology and Bioimaging (CENABIO), Rio de Janeiro, Brazil (GRID:grid.8536.8) (ISNI:0000 0001 2294 473X) 
 Federal University of Rio de Janeiro, School of Pharmacy, Rio de Janeiro, Brazil (GRID:grid.8536.8) (ISNI:0000 0001 2294 473X) 
Publication year
2022
Publication date
2022
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
2678586888
Copyright
© The Author(s) 2022. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.