A Novel Method to Reduce ELISA Serial Dilution Assay Workload Applied to SARS-CoV-2 and Seasonal HCoVs

  1. David Pattinson
  2. Peter Jester
  3. Lizheng Guan
  4. Seiya Yamayoshi
  5. Shiho Chiba
  6. Robert Presler
  7. Hongyu Rao
  8. Kiyoko Iwatsuki-Horimoto
  9. Nobuhiro Ikeda
  10. Masao Hagihara
  11. Tomoyuki Uchida
  12. Keiko Mitamura
  13. Peter Halfmann
  14. Gabriele Neumann
  15. Yoshihiro Kawaoka

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Abstract

Assays using ELISA measurements on serially diluted serum samples have been heavily used to measure serum reactivity to SARS-CoV-2 antigens and are widely used in virology and elsewhere in biology. We test a method using Bayesian hierarchical modelling to reduce the workload of these assays and measure reactivity of SARS-CoV-2 and HCoV antigens to human serum samples collected before and during the COVID-19 pandemic. Inflection titers for SARS-CoV-2 full-length spike protein (S1S2), spike protein receptor-binding domain (RBD), and nucleoprotein (N) inferred from 3 spread-out dilutions correlated with those inferred from 8 consecutive dilutions with an R2 value of 0.97 or higher. We confirm existing findings showing a small proportion of pre-pandemic human serum samples contain cross-reactive antibodies to SARS-CoV-2 S1S2 and N, and that SARS-CoV-2 infection increases serum reactivity to the beta-HCoVs OC43 and HKU1 S1S2. In serial dilution assays, large savings in resources and/or increases in throughput can be achieved by reducing the number of dilutions measured and using Bayesian hierarchical modelling to infer inflection or endpoint titers. We have released software for conducting these types of analysis.

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  1. Version published to 10.3390/v14030562
  2. ScreenIT

    SciScore for 10.1101/2021.09.13.21263523: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    NIH rigor criteria are not applicable to paper type.

    Table 2: Resources

    Antibodies
    SentencesResources
    Anti-human IgG-Peroxidase antibody produced in goat (Sera Care-KPL #.
    Anti-human IgG-Peroxidase
    suggested: None
    Microplates were reacted for 1 h at room temperature with the diluted serum samples, followed by peroxidase-conjugated goat anti-human IgG, Fcγ Fragment specific antibody (Jackson ImmunoResearch #109-035-098)
    anti-human IgG
    suggested: (Jackson ImmunoResearch Labs Cat# 109-035-098, RRID:AB_2337586)

    Results from OddPub: Thank you for sharing your code and data.

    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.

    Results from scite Reference Check: We found no unreliable references.

    About SciScore

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  3. Version published to 10.1101/2021.09.13.21263523v1 on medRxiv