Development of an ELISA for SARS-CoV-2 Detection focusing on antibodies against nucleocapsid protein

Document Type : Original Articles

Authors

1 Department of Proteomics and Biochemistry, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran

2 Razi Vaccine and Serum Research Institute

10.22092/ari.2024.366081.3188

Abstract

In late 2019, a new viral disease called COVID-19 caused by SARS-CoV-2 started in China and soon became a pandemic. This virus hurts human health and brings about financial damage to human societies, so researchers are trying to better identify and control it faster. Using the ELISA method to screen a large number of patients is always very helpful in infectious epidemics.
In this research, the nucleocapsid protein (NP) of the SARS-CoV-2 virus was used to measure serum antibodies, which was received from Avicenna Research Institute. We coated this antigen on each well of the plates of the ELISA test. After that, we added serum samples from medical diagnostic laboratories (positive and negative serums measured by ELISA and PCR. For this purpose, checkerboard titration for all serum samples and antigens was carried out to optimize ELISA. Our ELISA test was an indirect assay that could detect antibodies against nucleocapsid protein. Finally, we measured the cut-off, sensitivity, and specificity of our ELISA test.
Our results showed that 95% sensitivity and 92% specificity. Also, in the intra-assay, the value of CV was 0.263%and in the inter-assay, the value of CV was 0.41%. As a result, it can be stated that this ELISA test was highly accurate and had high durability.
In conclusion, our kit showcases promising capabilities in the accurate and efficient detection of antibodies targeted against NP. This innovative approach not only enhances diagnostic accuracy but also holds significant potential for advancing antibody detection methodologies in the field of virology and immunology.

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Archives of Razi Institute

Articles in Press, Accepted Manuscript
Available Online from 28 September 2024

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