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DataSheet_1_Development and evaluation of low-volume tests to detect and characterize antibodies to SARS-CoV-2.pdf

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posted on 2022-11-09, 13:31 authored by Alice Halliday, Anna E. Long, Holly E. Baum, Amy C. Thomas, Kathryn L. Shelley, Elizabeth Oliver, Kapil Gupta, Ore Francis, Maia Kavanagh Williamson, Natalie Di Bartolo, Matthew J. Randell, Yassin Ben-Khoud, Ilana Kelland, Georgina Mortimer, Olivia Ball, Charlie Plumptre, Kyla Chandler, Ulrike Obst, Massimiliano Secchi, Lorenzo Piemonti, Vito Lampasona, Joyce Smith, Michaela Gregorova, Lea Knezevic, Jane Metz, Rachael Barr, Begonia Morales-Aza, Jennifer Oliver, Lucy Collingwood, Benjamin Hitchings, Susan Ring, Linda Wooldridge, Laura Rivino, Nicholas Timpson, Jorgen McKernon, Peter Muir, Fergus Hamilton, David Arnold, Derek N. Woolfson, Anu Goenka, Andrew D. Davidson, Ashley M. Toye, Imre Berger, Mick Bailey, Kathleen M. Gillespie, Alistair J. K. Williams, Adam Finn

Low-volume antibody assays can be used to track SARS-CoV-2 infection rates in settings where active testing for virus is limited and remote sampling is optimal. We developed 12 ELISAs detecting total or antibody isotypes to SARS-CoV-2 nucleocapsid, spike protein or its receptor binding domain (RBD), 3 anti-RBD isotype specific luciferase immunoprecipitation system (LIPS) assays and a novel Spike-RBD bridging LIPS total-antibody assay. We utilized pre-pandemic (n=984) and confirmed/suspected recent COVID-19 sera taken pre-vaccination rollout in 2020 (n=269). Assays measuring total antibody discriminated best between pre-pandemic and COVID-19 sera and were selected for diagnostic evaluation. In the blind evaluation, two of these assays (Spike Pan ELISA and Spike-RBD Bridging LIPS assay) demonstrated >97% specificity and >92% sensitivity for samples from COVID-19 patients taken >21 days post symptom onset or PCR test. These assays offered better sensitivity for the detection of COVID-19 cases than a commercial assay which requires 100-fold larger serum volumes. This study demonstrates that low-volume in-house antibody assays can provide good diagnostic performance, and highlights the importance of using well-characterized samples and controls for all stages of assay development and evaluation. These cost-effective assays may be particularly useful for seroprevalence studies in low and middle-income countries.

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