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Article

Development of an Enzyme-Linked Immunosorbent Assay (ELISA) for Accurate and Prompt Coronavirus Disease 2019 (COVID-19) Diagnosis Using the Rational Selection of Serological Biomarkers

1
First Department of Paediatrics, “Aghia Sophia” Children’s Hospital, Immunology and Vaccinology Research Laboratory and Infectious Diseases Department “MAKKA”, Athens Medical School, 11527 Athens, Greece
2
University Research Institute for the Study of Genetic & Malignant Disorders in Childhood, First Department of Paediatrics, Aghia Sofia Children’s Hospital, Athens Medical School, 11527 Athens, Greece
3
Department of Microbiology, Athens Medical School, 11527 Athens, Greece
*
Author to whom correspondence should be addressed.
Academic Editor: Anna Baraniak
Diagnostics 2021, 11(11), 1970; https://doi.org/10.3390/diagnostics11111970
Received: 16 September 2021 / Revised: 20 October 2021 / Accepted: 20 October 2021 / Published: 23 October 2021
Prompt COVID-19 diagnosis is urgently required to support infection control measures. Currently available serological tests for measuring SARS-CoV-2 antibodies use different target antigens, although their sensitivity and specificity presents a challenge. We aimed to develop an “in-house” serological ELISA to measure antibodies against SARS-CoV-2 by combining different protein antigens. Sera (n = 44) from COVID-19-confirmed patients were evaluated against different SARS-CoV-2 protein antigens and all potential combinations using ELISA. Patients’ sera were also evaluated against commercially available ELISA diagnostic kits. The mixture containing RBD 2.5 μg/mL, S2 1 μg/mL and N 1.5 μg/mL was found to be the most potent. Plates were incubated with patients’ sera (1:100), and goat anti-human alkaline phosphatase-conjugated IgG, ΙgM and IgA antibody was added. The cut-off value for each assay was determined using the mean optical density plus two standard deviations of pre-pandemic controls. The “in-house” ELISA displayed 91% sensitivity and 97% specificity for IgG antibodies, whereas its sensitivity and specificity for IgM and IgA were 75% and 95% and 73% and 91%, respectively. The “in-house” ELISA developed here combined three SARS-CoV-2 antigens (RBD, S2 and N) as capture antigens and displayed comparable and even higher sensitivity and specificity than otherwise quite reliable commercially available ELISA diagnostic kits. View Full-Text
Keywords: SARS-CoV-2; antibodies; serological tests SARS-CoV-2; antibodies; serological tests
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MDPI and ACS Style

Lagousi, T.; Routsias, J.; Spoulou, V. Development of an Enzyme-Linked Immunosorbent Assay (ELISA) for Accurate and Prompt Coronavirus Disease 2019 (COVID-19) Diagnosis Using the Rational Selection of Serological Biomarkers. Diagnostics 2021, 11, 1970. https://doi.org/10.3390/diagnostics11111970

AMA Style

Lagousi T, Routsias J, Spoulou V. Development of an Enzyme-Linked Immunosorbent Assay (ELISA) for Accurate and Prompt Coronavirus Disease 2019 (COVID-19) Diagnosis Using the Rational Selection of Serological Biomarkers. Diagnostics. 2021; 11(11):1970. https://doi.org/10.3390/diagnostics11111970

Chicago/Turabian Style

Lagousi, Theano, John Routsias, and Vana Spoulou. 2021. "Development of an Enzyme-Linked Immunosorbent Assay (ELISA) for Accurate and Prompt Coronavirus Disease 2019 (COVID-19) Diagnosis Using the Rational Selection of Serological Biomarkers" Diagnostics 11, no. 11: 1970. https://doi.org/10.3390/diagnostics11111970

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