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A Highly Sensitive Immunoassay for Determination of Immune Response to SARS-CoV-2 in Capillary Blood Samples
Throughout the pandemic, serological assays have been revealed as crucial for detecting previous exposures to the virus and determining the timing of antibody maintenance after vaccination or natural infection. This study aimed to develop an optimized enzyme-linked immunosorbent assay (ELISA)-based...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9687217/ https://www.ncbi.nlm.nih.gov/pubmed/36428468 http://dx.doi.org/10.3390/biomedicines10112897 |
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author | Sánchez, Belén G. Bort, Alicia Mora-Rodríguez, José María Díaz-Yuste, Alba Gasalla, José Manuel Sánchez-Chapado, Manuel Sebastián-Martín, Alba Díaz-Laviada, Inés |
author_facet | Sánchez, Belén G. Bort, Alicia Mora-Rodríguez, José María Díaz-Yuste, Alba Gasalla, José Manuel Sánchez-Chapado, Manuel Sebastián-Martín, Alba Díaz-Laviada, Inés |
author_sort | Sánchez, Belén G. |
collection | PubMed |
description | Throughout the pandemic, serological assays have been revealed as crucial for detecting previous exposures to the virus and determining the timing of antibody maintenance after vaccination or natural infection. This study aimed to develop an optimized enzyme-linked immunosorbent assay (ELISA)-based serology, which could be used in case of reagent shortages, such as that occurred in the beginning of this health emergency. As a result, we present a high-sensitive immunoassay for the determination of IgG levels in venous serum samples, using 2 μg/mL antigen (receptor-binding domain of the spike protein S1) for coating the plate and utilizing human samples at a dilution 1:1000. This method showed non-inferiority features versus a commercial kit, is less expensive, and has a higher spectrophotometric range that allows for a better quantification of the antibody titers. The optical density values before and after heating venous serum samples at 56 °C during 30 min was quite similar, showing that heat inactivation can be used to reduce the biohazardous risks while handling samples. Furthermore, we show that finger-stick capillary blood samples can also serve as a suitable source for IgG detection, bypassing the need for serum isolation and being suitable for point-of-care application (Pearson’s coefficient correlation with capillary serum was 0.95, being statistically significant). |
format | Online Article Text |
id | pubmed-9687217 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-96872172022-11-25 A Highly Sensitive Immunoassay for Determination of Immune Response to SARS-CoV-2 in Capillary Blood Samples Sánchez, Belén G. Bort, Alicia Mora-Rodríguez, José María Díaz-Yuste, Alba Gasalla, José Manuel Sánchez-Chapado, Manuel Sebastián-Martín, Alba Díaz-Laviada, Inés Biomedicines Article Throughout the pandemic, serological assays have been revealed as crucial for detecting previous exposures to the virus and determining the timing of antibody maintenance after vaccination or natural infection. This study aimed to develop an optimized enzyme-linked immunosorbent assay (ELISA)-based serology, which could be used in case of reagent shortages, such as that occurred in the beginning of this health emergency. As a result, we present a high-sensitive immunoassay for the determination of IgG levels in venous serum samples, using 2 μg/mL antigen (receptor-binding domain of the spike protein S1) for coating the plate and utilizing human samples at a dilution 1:1000. This method showed non-inferiority features versus a commercial kit, is less expensive, and has a higher spectrophotometric range that allows for a better quantification of the antibody titers. The optical density values before and after heating venous serum samples at 56 °C during 30 min was quite similar, showing that heat inactivation can be used to reduce the biohazardous risks while handling samples. Furthermore, we show that finger-stick capillary blood samples can also serve as a suitable source for IgG detection, bypassing the need for serum isolation and being suitable for point-of-care application (Pearson’s coefficient correlation with capillary serum was 0.95, being statistically significant). MDPI 2022-11-11 /pmc/articles/PMC9687217/ /pubmed/36428468 http://dx.doi.org/10.3390/biomedicines10112897 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Sánchez, Belén G. Bort, Alicia Mora-Rodríguez, José María Díaz-Yuste, Alba Gasalla, José Manuel Sánchez-Chapado, Manuel Sebastián-Martín, Alba Díaz-Laviada, Inés A Highly Sensitive Immunoassay for Determination of Immune Response to SARS-CoV-2 in Capillary Blood Samples |
title | A Highly Sensitive Immunoassay for Determination of Immune Response to SARS-CoV-2 in Capillary Blood Samples |
title_full | A Highly Sensitive Immunoassay for Determination of Immune Response to SARS-CoV-2 in Capillary Blood Samples |
title_fullStr | A Highly Sensitive Immunoassay for Determination of Immune Response to SARS-CoV-2 in Capillary Blood Samples |
title_full_unstemmed | A Highly Sensitive Immunoassay for Determination of Immune Response to SARS-CoV-2 in Capillary Blood Samples |
title_short | A Highly Sensitive Immunoassay for Determination of Immune Response to SARS-CoV-2 in Capillary Blood Samples |
title_sort | highly sensitive immunoassay for determination of immune response to sars-cov-2 in capillary blood samples |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9687217/ https://www.ncbi.nlm.nih.gov/pubmed/36428468 http://dx.doi.org/10.3390/biomedicines10112897 |
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