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

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.

The relationship between IgG and IgM levels and severity of symptoms in COVID-19 patients confirmed by rapid antigen test.

This study aims to measure immunoglobulin G (IgG) and immunoglobulin M (IgM) response after detection of Severe Acute Respiratory Syndrome coronavirus (SARS CoV-2) antigens in coronavirus disease 2019 (COVID-19) patients concerning the severity of symptoms. SARS CoV-2 antigen was confirmed by rapid antigen test, and IgG and IgM were confirmed by VIDAS® SARS-COV-2 IgM and VIDAS® SARS-CoV-2 IgG automated qualitative assays used to rapidly detect antibodies 20-30 days after detection. The serological assay for detecting SARS-CoV-2 IgM and IgG antibodies shows a positive correlation for all patients detected with SARS-CoV-2 antigen with sensitivity 100% with differences in antibodies levels between patients regarding age and significantly related clinical symptoms with p-value 0.013 <0.05. The appearance of clinical symptoms was not significantly related to IgG levels at a p-value of 0.4 >0.05. However, the appearance of clinical symptoms was significantly related to IgM levels at a p-value of 0.002 <0.05. Antigen-dependent rapid tests can be used to detect SARS-CoV-2 in an early stage of infection with high sensitivity and specificity. Moreover, this study shows the age groups 21-30 and 31-40 have a better response to SARS-CoV-2 infection.