Sensitive and specific serological ELISA for the detection of SARS-CoV-2 infections

Luo, Ji; Brakel, Alexandra; Krizsan, Andor; Ludwig, Tobias; Mötzing, Marina; Volke, Daniela; Lakowa, Nicole; Grünewald, Thomas; Lehmann, Claudia; Wolf, Johannes; Borte, Stephan; Milkovska‐Stamenova, Sanja; Gabert, Jörg; Fingas, Felix; Scholz, Markus; Hoffmann, Ralf

doi:10.1186/s12985-022-01768-4

Cited by 23 publications

(28 citation statements)

References 33 publications

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“…Another advantage of using the N protein as an antigen is the possibility of differentiating infected and noninfected vaccinated persons producing anti-S-protein antibodies. Although our data are in accordance with reports in the literature, a limitation of our work was the use of a single antigen, not taking into account the possible variability that may exist in the protein production and purification procedure [34].…”

Section: Discussionsupporting

confidence: 86%

“…Noteworthy, previous studies regarding immunologic responses against the coronavirus subfamily indicated that the IgG antibody response against the N protein was more prominent than against the S protein [ 32 , 33 ]. In addition, previous studies with SARS-CoV-1 demonstrate that the serum durability of antibodies against the N protein is greater than antibodies against the S protein, making its use attractive for tracking viral infections for longer periods [ 34 , 35 ]. Another advantage of using the N protein as an antigen is the possibility of differentiating infected and noninfected vaccinated persons producing anti-S-protein antibodies.…”

Section: Discussionmentioning

confidence: 99%

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Diagnostic Performance of Three ELISAs for Detection of Antibodies against SARS-CoV-2 in Human Samples

Meira

Silva

Santos

et al. 2022

The Scientific World Journal

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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection that causes coronavirus disease 2019 (COVID-19) is a disease with a high rate of transmission. Serological tests are important to perform surveys and to determine the immunological status of the population. Based on this, we evaluated three enzyme-linked immunoassays (ELISAs) using different antigens from SARS-CoV-2 in a cohort of 161 patients. The performance of the ELISA developed for immunoglobulin G (IgG) measurement against SARS-CoV-2 was evaluated based on sensitivity, specificity, and accuracy. We found specificities of 0.98, 0.98, and 0.99 and sensitivities of 0.99, 0.91, and 0.87 for the nucleocapsid (N) protein, spike protein, and receptor binding domain (RBD) fraction, respectively. The accuracy assessment indicated the N protein (accuracy = 0.98) as the antigen most likely to give a correct diagnosis. Overall, the antibody responses were present for all three proteins in subjects with confirmed SARS-CoV-2 infections, showing a similar pattern of antibody production for different antigens. In summary, these highly sensitive and specific ELISAs, with a more competitive price, appear to be a valid approach for the serodiagnosis of COVID-19.

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Section: Discussionsupporting

confidence: 86%

Section: Discussionmentioning

confidence: 99%

Diagnostic Performance of Three ELISAs for Detection of Antibodies against SARS-CoV-2 in Human Samples

Meira

Silva

Santos

et al. 2022

The Scientific World Journal

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“…Proteins were purified by immobilized metal ion affinity chromatography (IMAC) using a HisTrap TM HP column (GE Healthcare, Chicago, IL, USA). The N protein was expressed as previously described [17].…”

Section: Recombinant Proteins Expressed In E Colimentioning

confidence: 99%

“…The cutoff value was determined by Receiver Operating Characteristics (ROC) analysis to obtain the best sensitivity and specificity based on a maximal Youden index using GraphPad Prism versions 9.0.2 or 5.01 (Graph Pad Software, La Jolla, CA, USA). N-protein ELISA was performed as previously described [17].…”

Section: Enzyme-linked Immunosorbent Assay (Elisa)mentioning

confidence: 99%

“…Most laboratories rely on commercial serological assays detecting anti-SARS-CoV-2 antibodies on S-and N-proteins. However, varied sensitivity and specificity were observed [11][12][13][14][15][16][17] and the sources of recombinantly expressed proteins were often not mentioned or the methods were not reported in detail. Here, we explored the potential of an in-house M-protein-based ELISA in comparison to in-house RBD-and N-protein-based ELISA for serological testing of SARS-CoV-2 infections.…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of S- and M-Proteins Expressed in Escherichia coli and HEK Cells for Serological Detection of Antibodies in Response to SARS-CoV-2 Infections and mRNA-Based Vaccinations

et al. 2022

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This study investigated the IgG and IgA antibody response against recombinant S1 and receptor binding domains (RBD) of the spike (S-) protein and the membrane (M-) protein using a set of 115 serum samples collected from patients infected with SARS-CoV-2 in Germany before April 2021 using protein and peptide ELISA. As S1- and RBD-proteins expressed in Escherichia coli provided poor sensitivities in ELISA, they were replaced by proteins expressed in HEK cells. The RBD-ELISA provided a sensitivity of 90.6% (N = 85) for samples collected from patients with confirmed SARS-CoV-2 infections more than 14 days after symptom onset or a positive PCR test. In population-based controls, the specificity was 97.9% (N = 94). In contrast, the sensitivities were only 41.2% and 72.6% for M- and N-proteins, respectively, while the specificities were 88.5% and 100%, respectively. Considering also 20 samples collected during the first two weeks of symptom onset or PCR confirmation, the sensitivity of RBD- and N-protein ELISA decreased to 82.6% and 72.6%, respectively. The combination of two data sets, i.e., N- and RBD-, N- and M-, or RBD- and M-proteins increased the sensitivity to 85.8%, 77.9%, and 87.8%, respectively. Peptide mapping mostly confirmed epitopes previously reported for S1- and M-proteins, but they were only recognized by a few samples already tested positive in the corresponding protein ELISA indicating that peptide-based assays will not improve the diagnostic sensitivity.

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Development of an Affordable ELISA Targeting the SARS-CoV-2 Nucleocapsid and Its Application to Samples from the Ongoing COVID-19 Epidemic in Ghana

Tapela,

Opurum,

Nuokpem

et al. 2023

Mol Diagn Ther

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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.

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Sensitive and specific serological ELISA for the detection of SARS-CoV-2 infections

Cited by 23 publications

References 33 publications

Diagnostic Performance of Three ELISAs for Detection of Antibodies against SARS-CoV-2 in Human Samples

Diagnostic Performance of Three ELISAs for Detection of Antibodies against SARS-CoV-2 in Human Samples

Evaluation of S- and M-Proteins Expressed in Escherichia coli and HEK Cells for Serological Detection of Antibodies in Response to SARS-CoV-2 Infections and mRNA-Based Vaccinations

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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