Development and Evaluation of ELISA Method for the Detection of SARS-CoV-2 RBD Protein Antibody
Keywords:
Covid-19, SARS-CoV-2, RBD, ELISA, antibodyAbstract
COVID-19 is caused by SARS-CoV-2 virus, an emerging virus classified as a highly pathogenic virus that has spread worldwide. The antibody against SARS-CoV-2 receptor binding domain (RBD) can be stimulated via natural infection or vaccination. In this study, the anti SARS-CoV-2 RBD IgG ELISA for the qualitative, semi-quantitative, and quantitative determination of infected and vaccinated human serum or plasma samples was developed. Analysis results of the qualitative detection revealed the cut-off values of the 1:200 dilution of negative sera, which were set up as mean values with two and three standard deviations, were 1.4447 and 1.8223 ELISA units and the specificity of negative sera were 93.94% and 98.27%, respectively. Furthermore, the sensitivity of positive sera at both cut-off values were 100%. The cross- reactivity were 6% and 0% at 1.4447 and 1.8232 cut-off values, respectively. There was no significant correlation between interferences and the sample sera (p > 0.05). For semi-quantitative determination, antibody concentration of 1:200 diluted samples could be determined via the standard curve ranging between 13.48-62.50 BAU/mL. However, higher antibody concentration sera had to be more diluted than 1:200 to be quantified by using the same standard curve. Additionally, when the antibody quantitative ELISA results were compared to the quantitative analysis results using chemiluminescence microparticle immunoassay (CMIA) the correlation of 0.93 (p < 0.001) was obtained. Consequently, the newly developed ELISA method can be further used to determine the antibody against SARS-CoV-2 RBD.
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