Evaluation of a Commercial Culture-Free Neutralization Antibody Detection Kit for Severe Acute Respiratory Syndrome-Related Coronavirus-2 and Comparison With an Antireceptor-Binding Domain Enzyme-Linked Immunosorbent Assay

  1. Jesse Papenburg
  2. Matthew P Cheng
  3. Rachel Corsini
  4. Chelsea Caya
  5. Emelissa Mendoza
  6. Kathy Manguiat
  7. L Robbin Lindsay
  8. Heidi Wood
  9. Michael A Drebot
  10. Antonia Dibernardo
  11. Gerasimos Zaharatos
  12. Reneé Bazin
  13. Romain Gasser
  14. Mehdi Benlarbi
  15. Gabrielle Gendron-Lepage
  16. Guillaume Beaudoin-Bussières
  17. Jérémie Prévost
  18. Andrés Finzi
  19. Momar Ndao
  20. Cedric P Yansouni

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Abstract

Background

Severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) surrogate neutralization assays that obviate the need for viral culture offer substantial advantages regarding throughput and cost. The cPass SARS-CoV-2 Neutralization Antibody Detection Kit (GenScript) is the first such commercially available assay that detects antibodies that block receptor-binding domain (RBD)/angiotensin-converting enzyme (ACE)-2 interaction. We aimed to evaluate cPass to inform its use and assess its added value compared with anti-RBD enzyme-linked immunosorbent assays (ELISAs).

Methods

Serum reference panels comprising 205 specimens were used to compare cPass to plaque-reduction neutralization test (PRNT) and a pseudotyped lentiviral neutralization (PLV) assay for detection of neutralizing antibodies. We assessed the correlation of cPass with an ELISA detecting anti-RBD immunoglobulin (Ig)G, IgM, and IgA antibodies at a single timepoint and across intervals from onset of symptoms of SARS-CoV-2 infection.

Results

Compared with PRNT-50, cPass sensitivity ranged from 77% to 100% and specificity was 95% to 100%. Sensitivity was also high compared with the pseudotyped lentiviral neutralization assay (93%; 95% confidence interval [CI], 85–97), but specificity was lower (58%; 95% CI, 48–67). Highest agreement between cPass and ELISA was for anti-RBD IgG (r = 0.823). Against the pseudotyped lentiviral neutralization assay, anti-RBD IgG sensitivity (99%; 95% CI, 94–100) was very similar to that of cPass, but overall specificity was lower (37%; 95% CI, 28–47). Against PRNT-50, results of cPass and anti-RBD IgG were nearly identical.

Conclusions

The added value of cPass compared with an IgG anti-RBD ELISA was modest.

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  1. Version published to 10.1093/ofid/ofab220
  2. ScreenIT

    SciScore for 10.1101/2021.01.23.21250325: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    In addition to panels using neutralization assays as the reference standard, we assembled 136 specimens from healthy blood donors who tested negative for the presence of anti-SARS-CoV-2 antibodies by both a lab-developed anti-RBD IgG ELISA and a commercial assay detecting anti-nucleocapsid antibodies (Abbott Architect SARS-CoV-2 IgG Assay).
    anti-SARS-CoV-2
    suggested: None
    anti-RBD IgG
    suggested: None
    anti-nucleocapsid
    suggested: None
    PRNT-50 titres and PRNT-90 titres ≥1:20 were considered positive for SARS-CoV-2 neutralizing antibodies.
    SARS-CoV-2 neutralizing antibodies.
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    After 1 hour of incubation at 37°C and 5% CO2, the sera-virus mixtures were added to 12-well plates containing Vero E6 cells at 90% to 100% confluence and incubated at 37°C and 5% CO2 for 1 hour.
    Vero E6
    suggested: None
    HEK 293T cells were transfected by the calcium phosphate method with the lentiviral vector pNL4.3 R-E- Luc (NIH AIDS Reagent Program) and a plasmid encoding for SARS- CoV-2 Spike at a ratio of 5:4.
    HEK 293T
    suggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)
    Software and Algorithms
    SentencesResources
    In addition to panels using neutralization assays as the reference standard, we assembled 136 specimens from healthy blood donors who tested negative for the presence of anti-SARS-CoV-2 antibodies by both a lab-developed anti-RBD IgG ELISA and a commercial assay detecting anti-nucleocapsid antibodies (Abbott Architect SARS-CoV-2 IgG Assay).
    Abbott Architect
    suggested: (Abbott ARCHITECT i1000sr System, RRID:SCR_019328)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    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.
    • No funding statement was detected.
    • No protocol registration statement was detected.

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  3. Version published to 10.1101/2021.01.23.21250325v2 on medRxiv
  4. Version published to 10.1101/2021.01.23.21250325v1 on medRxiv