The detection of IgG class antibodies against SARS-CoV-2 nucleocapsid protein by application of nanoparticles.

ABSTRACT

OBJECTIVE: Enzyme-linked immunosorbent assay (ELISA) is a widely used biochemical analytical method for the detection of a biomarker, through a specific antigen-antibody reaction. A common with ELISA is the amount of concrete biomarkers falling below the detection limit. Thus, the approach that will contribute to enhanced sensitivity of enzyme-linked immunosorbent assay is of great importance for medical practice. To address this issue, we used nanoparticles to improve the detection limit of traditional ELISA. MATERIALS AND METHODS: 80 samples were used, for which the presence of IgG antibodies against SARS-CoV-2 nucleocapsid protein were already determined qualitatively.  We tested the samples using an in vitro ELISA kit [SARS-CoV-2 IgG ELISA, COVG0949 (NovaTec, Leinfelden-Echterdingen, Germany)]. Additionally, we tested the same sample with the same ELISA kit but with the addition of 50 nm diameter citrate-capped silver nanoparticles. The reaction was performed, and data were calculated according to manufacturer guidelines. To measure ELISA results absorbance (optical density - OD) at 450 nm was read. RESULTS: Greater absorbance values have been revealed in case of silver nanoparticles application (66 cases, 82.5%, p<0.05). ELISA with application of nanoparticles classified 19 equivocal cases as positive and 3 equivocal ones as negative, 1 negative case as equivocal. CONCLUSIONS: Our findings suggest that nanoparticles can be used to improve the sensitivity of ELISA method and increase the detection limit. Thus, it is logical and desirable to enhance the sensitivity of ELISA method by application of nanoparticles; the approach is low cost and with a positive impact on accuracy.