Development and evaluation of low-volume tests to detect and characterise antibodies to SARS-CoV-2
Arnold David 1Berger Imre 2Hitchings Benjamin 3Halliday Alice 3Gillespie Kathleen M 4Secchi Massimiliano 5Lampasona Vito 5Gregorova Michaela 3Knezevic Lea 6Metz Jane 7Collingwood Lucy 8Rivino Laura 3McKernon Jorgen 9Muir Peter 9Goenka Anu 10Long Anna E 4Baum Holly E 11Shelley Kathryn L 12Oliver Elizabeth 3Gupta Kapil 13Williamson Maia Kavanagh 3di Bartolo Natalie 13Randell Matthew J 4Kelland Ilana 4Thomas Amy C 14Ben-Khoud Yassin 4Williams Alistair JK 4Timpson Nicholas 15Barr Rachael 10Smith Joyce 3Piemonti Lorenzo 5Mortimer Georgina 4Toye Ashley M 16Obst Ulrike 17Chandler Kyla 4Finn Adam 18Ring Susan 8Oliver Jennifer 8Francis Ore 6Ball Olivia 4Plumptre Charlie 3Davidson Andrew D. 3Bailey Mick 6Woolfson Derek N 19Morales-Aza Begonia 3Hamilton Fergus 20Wooldridge Linda6
作者信息
- 1. Academic Respiratory Unit, Bristol Medical School, University of Bristol
- 2. School of Chemistry, University of Bristol||School of Biochemistry, Biomedical Sciences Building, University Walk, University of Bristol||Bristol BioDesign Institute, University of Bristol, Life Sciences Building, Tyndall Avenue
- 3. School of Cellular and Molecular Medicine, University of Bristol
- 4. Diabetes and Metabolism, Bristol Medical School, University of Bristol
- 5. Diabetes Research Institute, IRCCS San Raffaele Scientific Institute
- 6. Bristol Veterinary School, University of Bristol
- 7. School of Cellular and Molecular Medicine, University of Bristol||Department of Paediatric Immunology and Infectious Diseases, Bristol Royal Hospital for Children||Academic Respiratory Unit, Bristol Medical School, University of Bristol
- 8. Population Health Sciences, Bristol Medical School, University of Bristol
- 9. National Infection Service, UK Health Security Agency, Southmead Hospital
- 10. School of Cellular and Molecular Medicine, University of Bristol||Department of Paediatric Immunology and Infectious Diseases, Bristol Royal Hospital for Children
- 11. School of Cellular and Molecular Medicine, University of Bristol||School of Chemistry, University of Bristol
- 12. School of Chemistry, University of Bristol||School of Biochemistry, Biomedical Sciences Building, University Walk, University of Bristol
- 13. School of Biochemistry, Biomedical Sciences Building, University Walk, University of Bristol
- 14. Bristol Veterinary School, University of Bristol||Population Health Sciences, Bristol Medical School, University of Bristol
- 15. Population Health Sciences, Bristol Medical School, University of Bristol||MRC Integrative Epidemiology Unit at University of Bristol
- 16. School of Biochemistry, Biomedical Sciences Building, University Walk, University of Bristol||Bristol BioDesign Institute, University of Bristol, Life Sciences Building, Tyndall Avenue||Bristol Institute of Transfusion Sciences, NHS Blood and Transplant Filton
- 17. School of Chemistry, University of Bristol
- 18. School of Cellular and Molecular Medicine, University of Bristol||Population Health Sciences, Bristol Medical School, University of Bristol||Department of Paediatric Immunology and Infectious Diseases, Bristol Royal Hospital for Children
- 19. School of Chemistry, University of Bristol||School of Biochemistry, Biomedical Sciences Building, University Walk, University of Bristol||Bristol Institute of Transfusion Sciences, NHS Blood and Transplant Filton
- 20. MRC Integrative Epidemiology Unit at University of Bristol||Academic Respiratory Unit, Bristol Medical School, University of Bristol
- 折叠
Abstract
Abstract
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 utilised 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-characterised 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.
FundingThis work was supported by multiple grants to AH, AL, OF, AT, IB awarded by the Elizabeth Blackwell Institute, and funded in part by the Wellcome Trust [Grant number 204813/Z/16/Z] with additional support from Bristol Alumni and Friends. AEL is funded by a Diabetes UK/JDRF RD Lawrence Fellowship (18/0005778 and 3-APF-2018-591-A-N). The DISCOVER study was supported by donations to Southmead Hospital Charity (Registered Charity Number: 1055900). The LOGIC study was funded by The Grand Appeal (The Official Bristol Children’s Hospital Charity; a registered charity in England and Wales (1043603)) through a grant awarded to AF & AG. ACT is supported by the Wellcome Trust (217509/Z/19/Z) and UKRI through the JUNIPER consortium MR/V038613/1 and CoMMinS study MR/V028545/1. The UK Medical Research Council and Wellcome (Grant ref: 217065/Z/19/Z) and the University of Bristol provide core support for ALSPAC. NJT is a Wellcome Trust Investigator (202802/Z/16/Z), is the PI of the Avon Longitudinal Study of Parents and Children (MRC & WT 217065/Z/19/Z), is supported by the University of Bristol NIHR Biomedical Research Centre (BRC-1215-2001), the MRC Integrative Epidemiology Unit (MC_UU_00011/1) and works within the CRUK Integrative Cancer Epidemiology Programme (C18281/A29019). LIPS assay development was supported by a joint grant from Diabetes UK/JDRF (20/0006217) to KMG. IB is supported by the Wellcome Trust (106115/Z/14/Z, 221708/Z/20/Z), the ERC (contr. nrs. 834631, 963992) and the EPSRC Impact Acceleration Account EP/R511663/1. We also acknowledge funding from BBSRC/EPSRC Synthetic Biology Research Centre (BB/L01386X/1, to NDB and AMT), NHS Blood and Transplant (WP15-05, to NDB and AMT), and the NIHR Blood and Transplant Research Unit in Red Cell Products (IS-BTU-1214-10032, to NDB and AMT). This publication is the work of the authors and Alice Halliday et al will serve as guarantors for the contents of this paper.
Conflict of interestAF is a member of the Joint Committee on Vaccination and Immunisation, the UK national immunisation technical advisory group and is chair of the WHO European regional technical advisory group of experts (ETAGE) on immunisation and ex officio a member of the WHO SAGE working group on COVID vaccines. He is investigator on studies and trials funded by Pfizer, Sanofi, Valneva, the Gates Foundation and the UK government. This manuscript presents independent research funded in part by the National Institute for Health Research (NIHR). The views expressed are those of the authors and not necessarily those of the NHS, the NIHR, or the Department of Health and Social Care.引用本文复制引用
Arnold David,Berger Imre,Hitchings Benjamin,Halliday Alice,Gillespie Kathleen M,Secchi Massimiliano,Lampasona Vito,Gregorova Michaela,Knezevic Lea,Metz Jane,Collingwood Lucy,Rivino Laura,McKernon Jorgen,Muir Peter,Goenka Anu,Long Anna E,Baum Holly E,Shelley Kathryn L,Oliver Elizabeth,Gupta Kapil,Williamson Maia Kavanagh,di Bartolo Natalie,Randell Matthew J,Kelland Ilana,Thomas Amy C,Ben-Khoud Yassin,Williams Alistair JK,Timpson Nicholas,Barr Rachael,Smith Joyce,Piemonti Lorenzo,Mortimer Georgina,Toye Ashley M,Obst Ulrike,Chandler Kyla,Finn Adam,Ring Susan,Oliver Jennifer,Francis Ore,Ball Olivia,Plumptre Charlie,Davidson Andrew D.,Bailey Mick,Woolfson Derek N,Morales-Aza Begonia,Hamilton Fergus,Wooldridge Linda.Development and evaluation of low-volume tests to detect and characterise antibodies to SARS-CoV-2[EB/OL].(2025-03-28)[2026-04-03].https://www.medrxiv.org/content/10.1101/2022.05.03.22274395.学科分类
医学研究方法/基础医学/预防医学
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