2023, Volume 16, Issue 6, pp 883 – 889

Development of a cost-effective quantitative in-house ELISA assay for screening anti-S1 IgG antibodies targeting SARS-CoV-2

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Authors and Affiliations

Corresponding Author: Abdul-Sattar AL-Saeedi Medical Microbiology Department, College of Medicine, Al-Nahrain University, Baghdad, Iraq Basic and Medical Sciences Branch, College of Nursing, University of Al-Ameed, Karbala, Iraq Email: abdulsattaraswed@gmail.com

Ahmed Sahib Abdulamir Medical Microbiology Department, College of Medicine, Al-Nahrain University, Baghdad, Iraq Email: ahmsah73@yahoo.com

Abstract

The RBD, S, and N proteins, the three main antigens of the SARS-CoV-2 virus, activate the host immune system and cause the formation of IgM and IgG antibodies. While IgM indicates an early, acute infection stage, IgG shows a past infection or persistent sickness. This study used an indirect ELISA assay that targets the S1 subunit of the SARS-CoV-2 S protein to create an in-house, qualitative serological test specific to COVID-19. A total of 60 serum samples were examined using ELISA for anti-SARS-CoV-2 IgG, and 50 of those results were positive. An additional 20 samples were taken from cases that occurred before the pandemic. For the in-house ELISA assay, a plasmid containing the gene coding for the S1 subunit was transformed into E. coli DH5ɑ bacterial cells and the protein was synthesized and purified. The purified protein was utilized to coat the ELISA plate, which was subsequently used to assess the levels of IgG among individuals with SARS-CoV-2 infection. The study found a significant association (p-value=0.01) between the in-house and the commercial anti-S1 subunit IgG antibodies kits. The in-house ELISA responded well, with a sensitivity and specificity of 75.0% and 88.89%, respectively. Furthermore, a library of SARS-CoV-2 recombinant S1 subunits was created by competent bacteria and may be employed for various tasks, such as creating diagnostic tools and scientific investigation. Overall, the in-house anti-SARS-CoV-2 human IgG-ELISA proved to be sensitive and specific for identifying IgG antibodies in patients exposed to SARS-CoV-2.

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About this article

PMC ID: 10478653
PubMed ID: 37675173
DOI: 10.25122/jml-2023-0047

Article Publishing Date (print): 6 2023
Available Online: 

Journal information

ISSN Printing: 1844-122X
ISSN Online: 1844-3117
Journal Title: Journal of Medicine and Life

Copyright License: Open Access

This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use and redistribution provided that the original author and source are credited.


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