A Dual-Antigen Enzyme-Linked Immunosorbent Assay Allows the Assessment of Severe Acute Respiratory Syndrome Coronavirus 2 Antibody Seroprevalence in a Low-Transmission Setting

  1. Sarah M Hicks
  2. Kai Pohl
  3. Teresa Neeman
  4. Hayley A McNamara
  5. Kate M Parsons
  6. Jin-shu He
  7. Sidra A Ali
  8. Samina Nazir
  9. Louise C Rowntree
  10. Thi H O Nguyen
  11. Katherine Kedzierska
  12. Denise L Doolan
  13. Carola G Vinuesa
  14. Matthew C Cook
  15. Nicholas Coatsworth
  16. Paul S Myles
  17. Florian Kurth
  18. Leif E Sander
  19. Graham J Mann
  20. Russell L Gruen
  21. Amee J George
  22. Elizabeth E Gardiner
  23. Ian A Cockburn
  24. SARS-CoV-2 Testing in Elective Surgery Collaborators

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Abstract

Estimates of seroprevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies have been hampered by inadequate assay sensitivity and specificity. Using an enzyme-linked immunosorbent assay–based approach that combines data about immunoglobulin G responses to both the nucleocapsid and spike receptor binding domain antigens, we show that excellent sensitivity and specificity can be achieved. We used this assay to assess the frequency of virus-specific antibodies in a cohort of elective surgery patients in Australia and estimated seroprevalence in Australia to be 0.28% (95% Confidence Interval, 0–1.15%). These data confirm the low level of transmission of SARS-CoV-2 in Australia before July 2020 and validate the specificity of our assay.

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  1. Version published to 10.1093/infdis/jiaa623
  2. ScreenIT

    SciScore for 10.1101/2020.09.09.20191031: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementConsent: Samples and ethics statement: Collection of blood from individuals pre-2020 was carried out after provision of informed consent, using procedures approved by the Human Research Ethics Committees (HREC) of the Australian National University (2016/317) and ACT Health (1.16.011 and 1.15.015).
    IRB: #H7886); ACT Health HREC (1.16.011): Charité Ethics Committee (EA2/066/20) [7].
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    After 1 h incubation at RT, wells were washed five times with PBS-T and incubated with 100 µL of horseradish peroxidase (HRP)-conjugated anti-human IgG, IgM or IgA antibodies diluted to the optimal concentration in 1% BSA (w/v) in PBS with 0.1% Tween-20 for 1 h at RT.
    anti-human IgG
    suggested: (LSBio (LifeSpan Cat# LS-C6444-30, RRID:AB_860431)
    IgA
    suggested: None
    Software and Algorithms
    SentencesResources
    ROC analysis and cutoffs were determined using GraphPad Prism 8 software.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    Results from OddPub: Thank you for sharing your code.

    Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:
    A key caveat of our study is that the positive controls used for assay validation are skewed to hospitalized individuals and thus we do not know with certainty the performance characteristics of the assays for asymptomatic cases who are known to have lower antibody levels [4, 11]. Moreover, a recent study has suggested that asymptomatic cases may not always seroconvert, though the assays used there had lower sensitivity than we report for our assay [5, 11]. Overall however, these data suggest that the low case number seen in Australia was reflective of low community transmission not inadequate testing. This is supported by the fact that the subsequent outbreak in Melbourne in July/August 2020 emerged from breaches of hotel quarantine of overseas travelers rather than undetected community transmission. A variety of assays of have been put forward for the assessment of seroprevalence of antibodies to SARS-CoV-2. Lateral flow devices were used in early studies, but these devices have insufficient sensitivity and specificity for use in low prevalence settings [12]. However, more recent studies using ELISA based assays with greater statistical rigor have overcome some of these issues and given reliable estimates of seroprevalence in higher-transmission areas such as the United States [3, 13, 14]. More recently, commercial electrochemiluminescence-based assays have been developed that offer high degrees of sensitivity and specificity as well as standardization [6, 15]. However, the...

    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.
    • Thank you for including a protocol registration statement.

    About SciScore

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  3. Version published to 10.1101/2020.09.09.20191031v1 on medRxiv