Performance and validation of an adaptable multiplex assay for detection of serologic response to SARS-CoV-2 infection or vaccination

Kenny, Grace; Negi, Riya; O’Reilly, Sophie; Garcia-Leon, Alejandro; Alalwan, Dana; Gaillard, Colette Marie; Saini, Gurvin; Inzitari, Rosana; Feeney, Eoin; Yousif, Obada; Cotter, Aoife G.; Barra, Eoghan de; Sadlier, Corinna; Crispie, Fiona; Doran, Peter; Gautier, Virginie; Mallon, Patrick

doi:10.1101/2022.05.16.22275163

Cited by 2 publications

(2 citation statements)

References 21 publications

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“…The All Ireland Infectious Diseases Cohort Study is a prospective, multicentric study, enrolling COVID-19 patients, as described before ( 44 , 45 ). The study was approved by local institutional review boards.…”

Section: Methodsmentioning

confidence: 99%

Association of COVID-19 mortality with serum selenium, zinc and copper: Six observational studies across Europe

et al. 2022

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IntroductionCertain trace elements are essential for life and affect immune system function, and their intake varies by region and population. Alterations in serum Se, Zn and Cu have been associated with COVID-19 mortality risk. We tested the hypothesis that a disease-specific decline occurs and correlates with mortality risk in different countries in Europe.MethodsSerum samples from 551 COVID-19 patients (including 87 non-survivors) who had participated in observational studies in Europe (Belgium, France, Germany, Ireland, Italy, and Poland) were analyzed for trace elements by total reflection X-ray fluorescence. A subset (n=2069) of the European EPIC study served as reference. Analyses were performed blinded to clinical data in one analytical laboratory.ResultsMedian levels of Se and Zn were lower than in EPIC, except for Zn in Italy. Non-survivors consistently had lower Se and Zn concentrations than survivors and displayed an elevated Cu/Zn ratio. Restricted cubic spline regression models revealed an inverse nonlinear association between Se or Zn and death, and a positive association between Cu/Zn ratio and death. With respect to patient age and sex, Se showed the highest predictive value for death (AUC=0.816), compared with Zn (0.782) or Cu (0.769).DiscussionThe data support the potential relevance of a decrease in serum Se and Zn for survival in COVID-19 across Europe. The observational study design cannot account for residual confounding and reverse causation, but supports the need for intervention trials in COVID-19 patients with severe Se and Zn deficiency to test the potential benefit of correcting their deficits for survival and convalescence.

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Section: Methodsmentioning

confidence: 99%

Association of COVID-19 mortality with serum selenium, zinc and copper: Six observational studies across Europe

et al. 2022

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“…Others may offer protection against Wild-type (WT) SARS-CoV-2, the strain against which the vaccine S is modelled, but not against immune-escape Variants of Concern (VOC), including Beta and Omicron, which have amino acid mutations in key antibody binding sites in the RBD (12) . For this reason, an antibody titre, the measure of total anti-SARS-CoV-2 IgGs against a certain target present in a sample (13), is not sufficient to infer a protective immune response. Instead, the functional capacity of an antibody population can be determined using a neutralisation test.…”

Section: Introductionmentioning

confidence: 99%

Development of a Novel Medium Throughput Flow-Cytometry Based Micro-Neutralisation Test for SARS-CoV-2 with Applications in Clinical Vaccine Trials and Antibody Screening

O’Reilly

Kenny

Alrawahneh

et al. 2023

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Quantifying neutralising capacity of circulating SARS-COV-2 antibodies is critical in evaluating protective humoral immune responses generated post-infection/post-vaccination. Here we describe a novel medium-throughput flow cytometry-based micro-neutralisation test to evaluate Neutralising Antibody (NAb) responses against live SARS-CoV-2 Wild Type and Variants of Concern (VOC) in convalescent/vaccinated populations. Flow Cytometry-Based Micro-Neutralisation Test (Micro-NT) was performed in 96-well plates using clinical isolates WT-B, WT-B.177.18 and/or VOCs Beta and Omicron. Plasma samples (All Ireland Infectious Diseases (AIID) Cohort) were serially diluted (8 points, half-log) from 1/20 and pre-incubated with SARS-CoV-2 (1h, 37°C). Virus-plasma mixture were added onto VERO E6/VERO E6 TMPRSS2 cells for 18h. Percentage infected cells was analysed by automated flow cytometry following trypsinisation, fixation and SARS-CoV-2 Nucleoprotein intracellular staining. Half-maximal Neutralisation Titres (NT50) were determined using four-parameter logistic regression. Our assay was compared to Plaque Reduction Neutralisation Test (PRNT) and validated against WHO anti-SARS-CoV-2 Immunoglobulin Standards. Using WHO Standards with low, medium or high anti-SARS-CoV-2 IgG, both Micro-NT and PRNT achieved comparable NT50 values. Micro-NT was found to be highly reproducible (inter-assay CV of 11.64%). Screening 190 convalescent samples and 11 COVID-19 naive controls (AIID cohort) we demonstrated that Micro-NT has broad dynamic range differentiating NT50s <1/20 to >1/5000. We could also characterise immune-escape VOC observing up to 10-fold reduction in NT50 against SARS-CoV-2 Beta variant. Our flow cytometry-based Micro-NT is a robust and reliable assay to quantify NAb titres, and has been selected as an endpoint in clinical trials. It has higher throughput (96 well format versus 12 well) and reduced infection time (18h vs 48-96h) compared to the gold standard PRNT.

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Performance and validation of an adaptable multiplex assay for detection of serologic response to SARS-CoV-2 infection or vaccination

Cited by 2 publications

References 21 publications

Association of COVID-19 mortality with serum selenium, zinc and copper: Six observational studies across Europe

Association of COVID-19 mortality with serum selenium, zinc and copper: Six observational studies across Europe

Development of a Novel Medium Throughput Flow-Cytometry Based Micro-Neutralisation Test for SARS-CoV-2 with Applications in Clinical Vaccine Trials and Antibody Screening

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