Development of an Affordable ELISA Targeting the SARS-CoV-2 Nucleocapsid and Its Application to Samples from the Ongoing COVID-19 Epidemic in Ghana
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Published:2023-07-18
Issue:5
Volume:27
Page:583-592
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ISSN:1177-1062
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Container-title:Molecular Diagnosis & Therapy
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language:en
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Short-container-title:Mol Diagn Ther
Author:
Tapela Kesego, Opurum Precious C., Nuokpem Franklin Y., Tetteh Becky, Siaw Godfred K., Humbert Maria V., Tawiah-Eshun Sylvia, Barakisu Anna Ibrahim, Asiedu Kwame, Arhin Samuel Kojo, Manu Aaron A., Appiedu-Addo Sekyibea N. A., Obbeng Louisa, Quansah Darius, Languon Sylvester, Anyigba Claudia, Dosoo Daniel, Edu Nelson K. O., Oduro-Mensah Daniel, Ampofo William, Tagoe Emmanuel, Quaye Osbourne, Donkor Irene Owusu, Akorli Jewelna, Aniweh Yaw, Christodoulides Myron, Mutungi Joe, Bediako Yaw, Rayner Julian C, Awandare Gordon A, McCormick Christopher J., Quashie Peter KojoORCID
Abstract
Abstract
Introduction
The true nature of the population spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in populations is often not fully known as most cases, particularly in Africa, are asymptomatic. Finding the true magnitude of SARS-CoV-2 spread is crucial to provide actionable data about the epidemiological progress of the disease for researchers and policymakers. This study developed and optimized an antibody enzyme-linked immunosorbent assay (ELISA) using recombinant nucleocapsid antigen expressed in-house using a simple bacterial expression system.
Methods
Nucleocapsid protein from SARS-CoV-2 was expressed and purified from Escherichia coli. Plasma samples used for the assay development were obtained from Ghanaian SARS-CoV-2 seropositive individuals during the pandemic, while seronegative controls were plasma samples collected from blood donors before the coronavirus disease 2019 (COVID-19) pandemic. Another set of seronegative controls was collected during the COVID-19 pandemic. Antibody detection and levels within the samples were validated using commercial kits and Luminex. Analyses were performed using GraphPad Prism, and the sensitivity, specificity and background cut-off were calculated.
Results and Discussion
This low-cost ELISA (£0.96/test) assay has a high prediction of 98.9%, and sensitivity and specificity of 97% and 99%, respectively. The assay was subsequently used to screen plasma from SARS-CoV-2 RT-PCR-positive Ghanaians. The assay showed no significant difference in nucleocapsid antibody levels between symptomatic and asymptomatic, with an increase of the levels over time. This is in line with our previous publication.
Conclusion
This study developed a low-cost and transferable assay that enables highly sensitive and specific detection of human anti-SARS-CoV-2 IgG antibodies. This assay can be modified to include additional antigens and used for continuous monitoring of sero-exposure to SARS-CoV-2 in West Africa.
Funder
Bill and Melinda Gates Foundation
Publisher
Springer Science and Business Media LLC
Subject
Pharmacology,Genetics,Molecular Medicine,General Medicine
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