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Development of robust, indigenous ELISA for detection of IgG antibodies against CoV-2 N and S proteins: mass screening
ABSTRACT: The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) pandemic has adversely affected humankind and caused millions of deaths globally since January 2020. Robust and quick serological tests such as antibody detection assays for SARS-CoV-2 provide relevant information and aid in...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9382608/ https://www.ncbi.nlm.nih.gov/pubmed/35976427 http://dx.doi.org/10.1007/s00253-022-12113-8 |
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author | Srivastava, Ashish Kumar Gupta, Avinash Chauhan, Deepika Meena, Ramesh Chand Sugadev, Ragumani Eslavath, Malleswara Rao Gupta, Harshita Karuna Singh, Sayar Singh, Yamini Tiwari, R. P. Kohli, Veena Varshney, Rajeev Ganju, Lilly |
author_facet | Srivastava, Ashish Kumar Gupta, Avinash Chauhan, Deepika Meena, Ramesh Chand Sugadev, Ragumani Eslavath, Malleswara Rao Gupta, Harshita Karuna Singh, Sayar Singh, Yamini Tiwari, R. P. Kohli, Veena Varshney, Rajeev Ganju, Lilly |
author_sort | Srivastava, Ashish Kumar |
collection | PubMed |
description | ABSTRACT: The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) pandemic has adversely affected humankind and caused millions of deaths globally since January 2020. Robust and quick serological tests such as antibody detection assays for SARS-CoV-2 provide relevant information and aid in the process of vaccine development and diagnostics, as well as in sero-epidemiological monitoring of antibody response to the virus. The receptor-binding domain (RBD) of spike and nucleocapsid protein are specific targets for detecting SARS-CoV-2 antibodies. Here, we present the development of a stable spike (S) and nucleocapsid (N) protein-based ELISA antibody detection test “CoroSuchak,” with 99% sensitivity, 98% specificity, cost-effective, and detection in a minimum time for serodiagnosis and mass screening of the population for antibodies against SARS-CoV-2. Blood samples were analyzed from 374 SARS-CoV-2 reverse transcription-polymerase chain reaction (RT-PCR) positive, 772 negative and asymptomatic, and 874 random groups of subjects. We found that the antibody titer was significantly higher (p < 0.0001) in infected and vaccinated group compared to the only vaccinated and only infected group. Using enzyme-linked immunosorbent assay (ELISA), we detected SARS-CoV-2 immunoglobulin G (IgG) antibodies in 118/123 (96%) infected individuals, 570/653 (87%) non-infected but vaccinated individuals, 231/237 (97%) individuals who were both infected and vaccinated, and 499/874 (57%) from randomly selected individuals from the first and second waves of the pandemic. Similarly in the third wave, 14/14 (100%) infected and 16/20 (80%) RT-PCR-negative but symptomatic subjects were detected. Thus, the highly sensitive and specific in-house developed ELISA antibody detection kit “CoroSuchak” is extremely useful to determine the seroprevalence of SARS-CoV-2 antibodies in the coronavirus-exposed population. KEY POINTS: •Indigenous kit using a combination of spike and nucleocapsid proteins and peptide sequences. •High sensitivity and specificity to detect variants. •Highly sensitive for mass screening. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00253-022-12113-8. |
format | Online Article Text |
id | pubmed-9382608 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-93826082022-08-17 Development of robust, indigenous ELISA for detection of IgG antibodies against CoV-2 N and S proteins: mass screening Srivastava, Ashish Kumar Gupta, Avinash Chauhan, Deepika Meena, Ramesh Chand Sugadev, Ragumani Eslavath, Malleswara Rao Gupta, Harshita Karuna Singh, Sayar Singh, Yamini Tiwari, R. P. Kohli, Veena Varshney, Rajeev Ganju, Lilly Appl Microbiol Biotechnol Methods and Protocols ABSTRACT: The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) pandemic has adversely affected humankind and caused millions of deaths globally since January 2020. Robust and quick serological tests such as antibody detection assays for SARS-CoV-2 provide relevant information and aid in the process of vaccine development and diagnostics, as well as in sero-epidemiological monitoring of antibody response to the virus. The receptor-binding domain (RBD) of spike and nucleocapsid protein are specific targets for detecting SARS-CoV-2 antibodies. Here, we present the development of a stable spike (S) and nucleocapsid (N) protein-based ELISA antibody detection test “CoroSuchak,” with 99% sensitivity, 98% specificity, cost-effective, and detection in a minimum time for serodiagnosis and mass screening of the population for antibodies against SARS-CoV-2. Blood samples were analyzed from 374 SARS-CoV-2 reverse transcription-polymerase chain reaction (RT-PCR) positive, 772 negative and asymptomatic, and 874 random groups of subjects. We found that the antibody titer was significantly higher (p < 0.0001) in infected and vaccinated group compared to the only vaccinated and only infected group. Using enzyme-linked immunosorbent assay (ELISA), we detected SARS-CoV-2 immunoglobulin G (IgG) antibodies in 118/123 (96%) infected individuals, 570/653 (87%) non-infected but vaccinated individuals, 231/237 (97%) individuals who were both infected and vaccinated, and 499/874 (57%) from randomly selected individuals from the first and second waves of the pandemic. Similarly in the third wave, 14/14 (100%) infected and 16/20 (80%) RT-PCR-negative but symptomatic subjects were detected. Thus, the highly sensitive and specific in-house developed ELISA antibody detection kit “CoroSuchak” is extremely useful to determine the seroprevalence of SARS-CoV-2 antibodies in the coronavirus-exposed population. KEY POINTS: •Indigenous kit using a combination of spike and nucleocapsid proteins and peptide sequences. •High sensitivity and specificity to detect variants. •Highly sensitive for mass screening. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00253-022-12113-8. Springer Berlin Heidelberg 2022-08-17 2022 /pmc/articles/PMC9382608/ /pubmed/35976427 http://dx.doi.org/10.1007/s00253-022-12113-8 Text en © The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2022, Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic. |
spellingShingle | Methods and Protocols Srivastava, Ashish Kumar Gupta, Avinash Chauhan, Deepika Meena, Ramesh Chand Sugadev, Ragumani Eslavath, Malleswara Rao Gupta, Harshita Karuna Singh, Sayar Singh, Yamini Tiwari, R. P. Kohli, Veena Varshney, Rajeev Ganju, Lilly Development of robust, indigenous ELISA for detection of IgG antibodies against CoV-2 N and S proteins: mass screening |
title | Development of robust, indigenous ELISA for detection of IgG antibodies against CoV-2 N and S proteins: mass screening |
title_full | Development of robust, indigenous ELISA for detection of IgG antibodies against CoV-2 N and S proteins: mass screening |
title_fullStr | Development of robust, indigenous ELISA for detection of IgG antibodies against CoV-2 N and S proteins: mass screening |
title_full_unstemmed | Development of robust, indigenous ELISA for detection of IgG antibodies against CoV-2 N and S proteins: mass screening |
title_short | Development of robust, indigenous ELISA for detection of IgG antibodies against CoV-2 N and S proteins: mass screening |
title_sort | development of robust, indigenous elisa for detection of igg antibodies against cov-2 n and s proteins: mass screening |
topic | Methods and Protocols |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9382608/ https://www.ncbi.nlm.nih.gov/pubmed/35976427 http://dx.doi.org/10.1007/s00253-022-12113-8 |
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