SARS-CoV-2 Seroprevalence in a Cohort of Asymptomatic RT-PCR Negative Croatian First League Football Players

  1. Adriana Vince
  2. Renata Zadro
  3. Zvonimir Sostar
  4. Suncanica Ljubin Sternak
  5. Jasmina Vranes
  6. Vedrana Skaro
  7. Petar Projic
  8. Vilim Molnar
  9. Vid Matisic
  10. Bruno Barsic
  11. Gordan Lauc
  12. Zvjezdana Lovric-Makaric
  13. Zoran Bahtijarevic
  14. Tomislav Vlahovic
  15. Sandra Sikic
  16. Ozren Polasek
  17. Dragan Primorac

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Abstract

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  1. ScreenIT

    SciScore for 10.1101/2020.10.30.20223230: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIRB: Ethical approval for the study was gained from the Ethics Committee of the Institute of Public Health “dr.
    Consent: All of the included patients gave a signed written consent to be included in the study prior to testing.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variableStudy Population: A total of 350 participants including all registered football players and club staff members of the Croatian First Football National League participated in this study, all of whom were male aged from 17 to 71 years.

    Table 2: Resources

    Antibodies
    SentencesResources
    Serology: For serologic testing in our cohort, we have decided to use the CE marked commercial ELISA assay (Anti-SARS-CoV-2 IgA, Anti-SARS-CoV-2 IgG S and Anti-SARS-CoV-2 IgG NCP, Euroimmun Medizinische Labordiagnostika AG, Lübeck, Germany) which detects IgA and IgG antibodies to the S1-domain of spike protein, as well as IgG antibodies to nucleocapsid antigen (NCP), which is the antigen with the strongest immune dominance in the coronavirus family.
    Anti-SARS-CoV-2 IgA
    suggested: None
    Anti-SARS-CoV-2 IgG
    suggested: None
    IgG
    suggested: None
    S1-domain of spike protein, as well as IgG antibodies to nucleocapsid antigen (NCP)
    suggested: None
    To exclude the acute Epstein-Barr virus (EBV) infection and the presence of heterophilic antibodies that could cause cross-reactivity in tested samples, serological analyses were performed to determine IgM and IgG class antibodies against Epstein-Barr virus capsid antigen (Anti-EBV-CA IgM and Anti-EBV-CA IgG, respectively) and IgG against EBV nuclear antigen 1 (Anti-EBNA-1 IgG) supplied by Euroimmun Medizinische Labordiagnostika AG, Lübeck, Germany.
    IgG class antibodies against Epstein-Barr virus capsid antigen (Anti-EBV-CA IgM
    suggested: None
    Anti-EBV-CA IgG
    suggested: None
    IgG against EBV nuclear antigen 1
    suggested: None
    Anti-EBNA-1 IgG
    suggested: None
    Software and Algorithms
    SentencesResources
    Amplification and detection of SARS-CoV-2 were performed using GeneFinder COVID-19 Plus RealAmp Kit (Osang Healthcare Co. Ltd., Anyang (Dongan), Gyeonggi, South Korea) on Cobas Z 480 Instrument (Roche Diagnostics GmbH, Mannheim, Germany) according to the manufacturer’s protocol.
    GeneFinder
    suggested: (GENEFINDER, RRID:SCR_009190)
    Statistical analysis: The analysis was performed using SAS 9.4 program, SAS Institute, Cary, North Carolina.
    SAS Institute
    suggested: (Statistical Analysis System, RRID:SCR_008567)

    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: We detected the following sentences addressing limitations in the study:
    Obviously, there is a limitation of ELISA IgA performance due to false-positivity in individuals without symptoms, most probably caused by cross-reactivity to other human coronaviruses. However, all of the aforementioned studies have investigated only single serum samples from presumably negative individuals or COVID-19 patients, so no dynamics in antibody response could be observed. On the contrary, we have analyzed paired sera and found substantial positive IgA dynamics in 15 out of 305 (5%) of participants (Table 5, Figure 1). Borderline IgA results in asymptomatic patients are not easy to interpret, therefore we decided to leave out of conclusion the participants that had borderline results without any dynamics or presence of IgG antibodies simultaneously. IgG (S1) antibodies were detected in 6 cases only. Meyer and coworkers suggest that higher IgG (S1) cut-off value for seropositivity is needed (2.5) to secure an optimal specificity and positive predictive value (28). However, IgG (S1) was detectable at baseline only in 6 sera in our cohort, out of which 4 were borderline and 2 had positive OD ratios (1.37 and 4.25). IgG (S1) positivity was not observed in paired sera at follow up 2 months later. Ortho-Heller and coworkers found a stronger decrease for IgG (NCP) than for S1 specific antibodies looking at the longitudinal kinetics in the cohort of 20 non-hospitalized patients. They conclude that a single SARS-COV-2 antibody test should not be used to exclude or confirm a...

    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.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  2. Version published to 10.26502/fjppr.046
  3. Version published to 10.1101/2020.10.30.20223230v1 on medRxiv