Supplementary appendix

This appendix formed part of the original submission. We post it as supplied by the
authors.

Supplement to: Roarty C, Tonry C, McFetridge L, Mitchell H, Watson C, Waterfield T, on

behalf of the Covid Warriors research team. Kinetics and seroprevalence of SARS-CoV-2
antibodies in children. Lancet Infect Dis 2020; published online Nov 19. https://doi.
org/10.1016/S1473-3099(20)30884-7.
Appendix for: Kinetics and seroprevalence of SARS-CoV-2 antibodies in children

Methods Summary:
The full protocol has undergone peer-review and has been accepted for publication in BMJ
Open. A brief summary of the methods is given below:

Design
This multicentre observational prospective cohort study was designed to determine the
seroprevalence of SARS-CoV-2 antibodies in healthy children. This study protocol was
written in conjunction with the Strengthening the Reporting of Observational Studies in
Epidemiology (STROBE) guidelines.

Objectives
1. Report the seroprevalence of SARS-CoV-2 immunoglobulin M (IgM) and/or
immunoglobulin G (IgG) antibodies in healthy children aged 2-15 years old at baseline 2
months after enrolment and 6 months after enrolment.
2. Determine if antibodies persist at 2 months and 6 months.
3. Report the symptoms associated with COVID-19 in children.
Methods and Analysis

Setting
We aimed to recruit1000 participants from five centres (Belfast, Cardiff, Glasgow, London
and Manchester) between May and July 2020.

Participants
Children of healthcare workers who were aged between 2 and 15 years old at the time of
recruitment were eligible for inclusion. Children receiving antibiotics, admitted to hospital
within seven days of recruitment, receiving oral immunosuppressive treatment at the time of
recruitment or ever diagnosed with a malignancy were excluded.

Consent
Informed consent was obtained prior to inclusion including assent from the child. The
parent/child was free to decline/withdraw consent at any time without providing a reason and
without being subject to any resulting detriment.

Assessments and procedures

All participants underwent phlebotomy at baseline and at 2 months with further phlebotomy
planned 6 months after enrolment.

Data Collection
Study data were collected and managed using REDCap (Research Electronic Data Capture)
electronic data capture tools. Participants and their parents provided information relating to
illness episodes, suspected household exposure to SARS-CoV-2 and the outcome of any
molecular testing at each clinic appointment.

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Outcome Measures
Primary outcome measure:
• Presence of IgG and/or IgM antibodies to SARS-CoV-2 in serum plasma
Secondary outcome measures:
• SARS-CoV-2 infection confirmed by RT-QPCR testing of oral/nasal swabs and/or
crevicular fluid.

Statistical analysis plan

The study population were described in terms of demographic characteristics with sex,
median age and healthcare role of parents. Simple descriptive statistics (total number and
proportion) were used to describe symptomology, vaccination status and household contacts
and seroprevalence of SARS-CoV-2 antibodies. Logistic regression modelling was used to
estimate the probability of the presence of SARS-CoV-2 antibodies, adjusting for factors
including demographic features (age, gender) and symptomology (such as fever, cough,
fatigue), that are deemed statistically significant.

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Table: Summary of seroprevalence by site. (Number and % unless stated)

SITE FIRST ROUND SECOND ROUND FIRST ROUND SECOND ROUND

(NUMBER (NUMBER PERCENTAGE PERCENTAGE
REACTIVE) REACTIVE) REACTIVE (95% REACTIVE (95%CI)
CI)

ALL 992(68) 849(65) 6.9(5.4,8.6) 7.7(6.1,9.6)

BELFAST 215(2) 200(4) 0.9(0.2,3.3) 2(0.8,5.0)
CARDIFF 178(10) 134(7) 5.6(3.1,10.0) 5.2(2.6, 10.4)
GLASGOW 224(20) 210(19) 8.9(5.9,13.4) 9.1(5.9, 13.7)
LONDON 199(23) 135(20) 11.6(7.8,16.8) 14.8(9.8,21.8)
MANCHESTER 176(13) 170(15) 7.4(4.4,12.) 9.8(5.4,14.1)

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Declarations

• Registration - https://www.clinicaltrials.gov (trial registration: NCT0434740) on the

15/04/2020.
• Ethical approval was obtained from the London - Chelsea Research Ethics Committee
(REC Reference - 20/HRA/1731) and the Belfast Health & Social Care Trust
Research Governance (Reference 19147TW-SW).
• Conflicts of interests: None declared.
• Funding: This work was supported by HSC R&D Division, Public Health Agency
Ref: COM/5596/20. This funding source had no role in the design of this study and
will not have any role during its execution, analyses, interpretation of the data, or
decision to submit result.
• Authors contributions: Dr Waterfield is the chief investigator of the study and co-
ordinated the running of the study including data management and site training. Dr
Watson, Dr Tonry and Mr Roarty were involved with data collection, data
management, data analysis, sample processing and antibody testing. Dr McFetridge
and Dr Mitchell provided statistical expertise and performed the statistical analysis.
All authors contributed to the writing of the manuscript.
• Acknowledgements: We thank all of the children and their families who participated
in this study. We also thank all of the sites (Belfast Health and Social Care Trust, The
Ulster Independent Clinic, Cardiff and Vale University Health Board, NHS Greater
Glasgow and Clyde, Public Health England, London, Manchester University NHS
Foundation Trust, NIHR Manchester Clinical Research Facility) and staff who
participated in screening and enrolment.
• Data Sharing: All of the individual participant data collected during this study will be
available (including data dictionaries) on the Queen’s University Belfast database
within 3 months of completion of the study.

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The Covid Warriors research team

Elizabeth Waxman, Derek Fairley, Gala Roew-Setz, James McKenna, Peter Mallet, Ryan

Chrisite, Katherine Christie, Emma McManus, Christian Bennison, Kate Mullan and

Aleksandra Metryka, Rebecca Moore, Kathryn Ferris, Alison Watt, Claire McGinn, Steven

Foster, Jennifer Evans, Mark D Lyttle, Shazad Ahmad, Shamez Ladhani, Michael Corr, Julie-

Ann Maney, and Sharon Christie.