Validation of a SARS-CoV-2 spike protein ELISA for use in contact investigations and sero-surveillance
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Abstract
Since emergence of SARS-CoV-2 in late 2019, there has been a critical need to understand prevalence, transmission patterns, to calculate the burden of disease and case fatality rates. Molecular diagnostics, the gold standard for identifying viremic cases, are not ideal for determining true case counts and rates of asymptomatic infection. Serological detection of SARS-CoV-2 specific antibodies can contribute to filling these knowledge gaps. In this study, we describe optimization and validation of a SARS-CoV-2-specific-enzyme linked immunosorbent assay (ELISA) using the prefusion-stabilized form of the spike protein [1]. We performed receiver operator characteristic (ROC) analyses to define the specificities and sensitivities of the optimized assay and examined cross reactivity with immune sera from persons confirmed to have had infections with other coronaviruses. These assays will be used to perform contact investigations and to conduct large-scale, cross sectional surveillance to define disease burden in the population.
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SciScore for 10.1101/2020.04.24.057323: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources HRP-conjugated goat anti-human antibodies (KPL) diluted at 1:2000 in serum diluent were added to washed plates and incubated at 37°C in a humidified chamber for 1 hour. anti-human antibodies (KPLsuggested: (SeraCare KPL Cat# 5210-0158, RRID:AB_2773725)Experimental Models: Cell Lines Sentences Resources ELISA: The pre-fusion stabilized ectodomain of SARS-CoV-2 spike (S) was expressed in suspension adapted HEK-293 cells as previously described [1]. HEK-293suggest…SciScore for 10.1101/2020.04.24.057323: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources HRP-conjugated goat anti-human antibodies (KPL) diluted at 1:2000 in serum diluent were added to washed plates and incubated at 37°C in a humidified chamber for 1 hour. anti-human antibodies (KPLsuggested: (SeraCare KPL Cat# 5210-0158, RRID:AB_2773725)Experimental Models: Cell Lines Sentences Resources ELISA: The pre-fusion stabilized ectodomain of SARS-CoV-2 spike (S) was expressed in suspension adapted HEK-293 cells as previously described [1]. HEK-293suggested: CLS Cat# 300192/p777_HEK293, RRID:CVCL_0045)Software and Algorithms Sentences Resources Statistics were performed using GraphPad Prism software (7.04). GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
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