Comparative Study

. 2021 Dec 7;16(12):e0261003.

doi: 10.1371/journal.pone.0261003. eCollection 2021.

Dried blood spot specimens for SARS-CoV-2 antibody testing: A multi-site, multi-assay comparison

François Cholette^{1

2}, Christine Mesa¹, Angela Harris¹, Hannah Ellis¹, Karla Cachero¹, Philip Lacap¹, Yannick Galipeau³, Marc-André Langlois^{3

4}, Anne-Claude Gingras^{5

6}, Cedric P Yansouni⁷, Jesse Papenburg^{7

8}, Matthew P Cheng^{7

8}, Pranesh Chakraborty⁹, Derek R Stein^{2

10}, Paul Van Caeseele^{2

10

11

12}, Sofia Bartlett^{13

14}, Mel Krajden^{13

14}, David Goldfarb¹⁴, Allison McGeer^{12

15}, Carla Osiowy¹, Catherine Hankins^{12

16}, Bruce Mazer¹⁷, Michael Drebot¹, John Kim¹; COVID-19 Immunity Task Force (CITF) working group

Affiliations

¹ National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Canada.
² Department of Medical Microbiology and Infectious Diseases, University of Manitoba, Winnipeg, Canada.
³ Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, Canada.
⁴ The Centre for Infection, Immunity, and Inflammation (CI3), University of Ottawa, Ottawa, Canada.
⁵ Department of Molecular Genetics, University of Toronto, Toronto, Canada.
⁶ Lunenfeld-Tanenbaum Research Institute at Mount Sinai Hospital, Sinai Health System, Toronto, Canada.
⁷ McGill Interdisciplinary Initiative in Infection and Immunity, McGill University, Montréal, Canada.
⁸ Division of Microbiology, Department of Clinical Laboratory Medicine, Optilab Montréal-McGill University Health Centre, Montréal, Canada.
⁹ Newborn Screening Ontario, Children's Hospital of Eastern Ontario, Ottawa, Canada.
¹⁰ Cadham Provincial Laboratory, Manitoba Health, Winnipeg, Canada.
¹¹ Department of Pediatrics & Child Health, University of Manitoba, Winnipeg, Canada.
¹² COVID-19 Immunity Task Force, Montréal, Canada.
¹³ British Columbia Centre for Disease Control, Vancouver, Canada.
¹⁴ Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, Canada.
¹⁵ Division of Infectious Diseases, University of Toronto, Toronto, Canada.
¹⁶ Department of Epidemiology, Biostatistics, and Occupational Health, McGill University, Montréal, Canada.
¹⁷ Department of Pediatrics, McGill University, Montréal, Canada.

PMID: 34874948
PMCID: PMC8651133
DOI: 10.1371/journal.pone.0261003

Comparative Study

Dried blood spot specimens for SARS-CoV-2 antibody testing: A multi-site, multi-assay comparison

François Cholette et al. PLoS One. 2021.

. 2021 Dec 7;16(12):e0261003.

doi: 10.1371/journal.pone.0261003. eCollection 2021.

Authors

Affiliations

¹ National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Canada.
² Department of Medical Microbiology and Infectious Diseases, University of Manitoba, Winnipeg, Canada.
³ Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, Canada.
⁴ The Centre for Infection, Immunity, and Inflammation (CI3), University of Ottawa, Ottawa, Canada.
⁵ Department of Molecular Genetics, University of Toronto, Toronto, Canada.
⁶ Lunenfeld-Tanenbaum Research Institute at Mount Sinai Hospital, Sinai Health System, Toronto, Canada.
⁷ McGill Interdisciplinary Initiative in Infection and Immunity, McGill University, Montréal, Canada.
⁸ Division of Microbiology, Department of Clinical Laboratory Medicine, Optilab Montréal-McGill University Health Centre, Montréal, Canada.
⁹ Newborn Screening Ontario, Children's Hospital of Eastern Ontario, Ottawa, Canada.
¹⁰ Cadham Provincial Laboratory, Manitoba Health, Winnipeg, Canada.
¹¹ Department of Pediatrics & Child Health, University of Manitoba, Winnipeg, Canada.
¹² COVID-19 Immunity Task Force, Montréal, Canada.
¹³ British Columbia Centre for Disease Control, Vancouver, Canada.
¹⁴ Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, Canada.
¹⁵ Division of Infectious Diseases, University of Toronto, Toronto, Canada.
¹⁶ Department of Epidemiology, Biostatistics, and Occupational Health, McGill University, Montréal, Canada.
¹⁷ Department of Pediatrics, McGill University, Montréal, Canada.

PMID: 34874948
PMCID: PMC8651133
DOI: 10.1371/journal.pone.0261003

Abstract

The true severity of infection due to COVID-19 is under-represented because it is based on only those who are tested. Although nucleic acid amplifications tests (NAAT) are the gold standard for COVID-19 diagnostic testing, serological assays provide better population-level SARS-CoV-2 prevalence estimates. Implementing large sero-surveys present several logistical challenges within Canada due its unique geography including rural and remote communities. Dried blood spot (DBS) sampling is a practical solution but comparative performance data on SARS-CoV-2 serological tests using DBS is currently lacking. Here we present test performance data from a well-characterized SARS-CoV-2 DBS panel sent to laboratories across Canada representing 10 commercial and 2 in-house developed tests for SARS-CoV-2 antibodies. Three commercial assays identified all positive and negative DBS correctly corresponding to a sensitivity, specificity, positive predictive value, and negative predictive value of 100% (95% CI = 72.2, 100). Two in-house assays also performed equally well. In contrast, several commercial assays could not achieve a sensitivity greater than 40% or a negative predictive value greater than 60%. Our findings represent the foundation for future validation studies on DBS specimens that will play a central role in strengthening Canada's public health policy in response to COVID-19.

PubMed Disclaimer

Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

**Fig 1. Positive predictive values (PPV) and negative predictive values (NPV) by prevalence for each commercial and in-house assay on dried blood spot specimens.**
PPV and NPV is presented for *n =* 10 SARS-CoV-2 antibody negative DBS specimens and n = 10 SARS-CoV-2 antibody positive DBS specimens. One 6 mm (1/4 inch) punch was used for the EUROIMMUN assay and two 6 mm (1/4 inch) punches were used for the Platelia and in-house assays. Data from four 6 mm (1/4 inch) punches is presented for all other assays. EUROIMMUN = Anti-SARS-CoV-2 ELISA assay (EUROIMMUN, Lübeck, Germany). Platelia = SARS-CoV-2 assay (Bio-Rad, Hercules, California). LIASON = SARS-CoV-2 assay (DiaSorin, Saluggia, Italy). COV2G = SARS-CoV-2 COV2G assay (Siemens, Erlangen, Germany). COV2T = SARS-CoV-2 COV2T assay (Siemens). Elecsys S = Quantitative Anti-SARS-CoV-2 assay (Elecsys spike; Roche, Basel, Switzerland). Elecsys N = Anti-SARS-CoV-2 assay (Elecsys nucleocapsid; Roche). VITROS = Anti-SARS-CoV-2 assay (Ortho Clinical Diagnostics, Raritan, New Jersey). Architect = SARS-CoV-2 assay (Abbott, Mississauga, Canada). GSP/DELFIA = Anti-SARS-CoV-2 assay (PerkinElmer, Waltham, Massachusetts). In-house S (U of T) = In-house spike assay (University of Toronto). In-house RBD (U of T) = In-house RBD assay (University of Toronto). In-house N (U of T) = In-house nucleocapsid assay (University of Toronto). In-house S, mono (U of O) = In-house monoclonal spike assay (University of Ottawa). In-house RBD, mono (U of O) = In-house monoclonal RBD assay (University of Ottawa). In-house N, mono (U of O) = In-house monoclonal nucleocapsid assay (University of Ottawa). The VITROS assay could not achieve a specificity greater than 0% therefore, only PPV is shown.

**Fig 2**
Inter-assay concordance (A) and Cohen’s kappa coefficient (B) for SARS-CoV-2 antibody testing on dried blood spot specimens. Concordance (%) and Cohen’s kappa coefficient are presented for n = 10 SARS-CoV-2 antibody negative DBS specimens and n = 10 SARS-CoV-2 antibody positive DBS specimens. One 6 mm (1/4 inch) punch was used for the EUROIMMUN assay and two 6 mm (1/4 inch) punches were used for the Platelia and in-house assays. EUROIMMUN = Anti-SARS-CoV-2 ELISA assay (EUROIMMUN, Lübeck, Germany). Platelia = SARS-CoV-2 assay (Bio-Rad, Hercules, California). LIASON = SARS-CoV-2 assay (DiaSorin, Saluggia, Italy). COV2G = SARS-CoV-2 COV2G assay (Siemens, Erlangen, Germany). COV2T = SARS-CoV-2 COV2T assay (Siemens). Elecsys S = Quantitative Anti-SARS-CoV-2 assay (Elecsys spike; Roche, Basel, Switzerland). Elecsys N = Anti-SARS-CoV-2 assay (Elecsys nucleocapsid; Roche). VITROS = Anti-SARS-CoV-2 assay (Ortho Clinical Diagnostics, Raritan, New Jersey). Architect = SARS-CoV-2 assay (Abbott, Mississauga, Canada). GSP/DELFIA = Anti-SARS-CoV-2 assay (PerkinElmer, Waltham, Massachusetts). In-house S (U of T) = In-house spike assay (University of Toronto). In-house RBD (U of T) = In-house RBD assay (University of Toronto). In-house N (U of T) = In-house nucleocapsid assay (University of Toronto). In-house S, mono (U of O) = In- house monoclonal spike assay (University of Ottawa). In-house RBD, mono (U of O) = In-house monoclonal RBD assay (University of Ottawa). In-house N, mono (U of O) = In-house monoclonal nucleocapsid assay (University of Ottawa).

**Fig 3. Receiver operating characteristic (ROC) curve for each commercial and in-house assay on dried blood spot specimens.**
ROC curves are presented for n = 10 SARS-CoV-2 antibody negative DBS specimens and n = 10 SARS-CoV-2 antibody positive DBS specimens. One 6 mm (1/4 inch) punch was used for the EUROIMMUN assay and two 6 mm (1/4 inch) punches were used for the Platelia and in-house assays. Data from four 6 mm (1/4 inch) punches is presented for all other assays. EUROIMMUN = Anti-SARS-CoV-2 ELISA assay (EUROIMMUN, Lübeck, Germany). Platelia = SARS-CoV-2 assay (Bio-Rad, Hercules, California). LIASON = SARS-CoV-2 assay (DiaSorin, Saluggia, Italy). COV2G = SARS-CoV-2 COV2G assay (Siemens, Erlangen, Germany). COV2T = SARS-CoV-2 COV2T assay (Siemens). Elecsys S = Quantitative Anti-SARS-CoV-2 assay (Elecsys spike; Roche, Basel, Switzerland). Elecsys N = Anti-SARS-CoV-2 assay (Elecsys nucleocapsid; Roche). VITROS = Anti-SARS-CoV-2 assay (Ortho Clinical Diagnostics, Raritan, New Jersey). Architect = SARS-CoV-2 assay (Abbott, Mississauga, Canada). GSP/DELFIA = Anti-SARS-CoV-2 assay (PerkinElmer, Waltham, Massachusetts). In-house S (U of T) = In-house spike assay (University of Toronto). In-house RBD (U of T) = In-house RBD assay (University of Toronto). In-house N (U of T) = In-house nucleocapsid assay (University of Toronto). In-house S, mono (U of O) = In-house monoclonal spike assay (University of Ottawa). In-house RBD, mono (U of O) = In-house monoclonal RBD assay (University of Ottawa). In-house N, mono (U of O) = In-house monoclonal nucleocapsid assay (University of Ottawa).

**Fig 4. Distribution of values obtained for each commercial and in-house assay on dried blood spot (DBS) specimens.**
Each panel shows the optical density ratio (OD Ratio), arbitrary units per mL (AU/mL), index, units per mL (U/mL), cut-off index, or signal to cut-off ratio (S/Co) for SARS-CoV-2 antibody negative DBS specimens (n = 10) represented in blue and SARS-CoV-2 antibody positive DBS specimens (n = 10) represented in orange. All values are log₁₀ transformed to aid with visualisation. One 6 mm (1/4 inch) punch was used for the EUROIMMUN assay and two 6 mm (1/4 inch) punches were used for the Platelia and in-house assays. Data from four 6 mm (1/4 inch) punches is presented for all other assays. EUROIMMUN = Anti-SARS-CoV-2 ELISA assay (EUROIMMUN, Lübeck, Germany). Platelia = SARS-CoV-2 assay (Bio-Rad, Hercules, California). LIASON = SARS-CoV-2 assay (DiaSorin, Saluggia, Italy). COV2G = SARS-CoV-2 COV2G assay (Siemens, Erlangen, Germany). COV2T = SARS-CoV-2 COV2T assay (Siemens). Elecsys S = Quantitative Anti-SARS-CoV-2 assay (Elecsys spike; Roche, Basel, Switzerland). Elecsys N = Anti-SARS-CoV-2 assay (Elecsys nucleocapsid; Roche). VITROS = Anti-SARS-CoV-2 assay (Ortho Clinical Diagnostics, Raritan, New Jersey). Architect = SARS-CoV-2 assay (Abbott, Mississauga, Canada). GSP/DELFIA = Anti-SARS-CoV-2 assay (PerkinElmer, Waltham, Massachusetts). In-house S (U of T) = In-house spike assay (University of Toronto). In-house RBD (U of T) = In-house RBD assay (University of Toronto). In-house N (U of T) = In-house nucleocapsid assay (University of Toronto). In-house S, mono (U of O) = In-house monoclonal spike assay (University of Ottawa). In-house RBD, mono (U of O) = In-house monoclonal RBD assay (University of Ottawa). In-house N, mono (U of O) = In-house monoclonal nucleocapsid assay (University of Ottawa).

See this image and copyright information in PMC

Cited by

A bead-based multiplex assay covering all coronaviruses pathogenic for humans for sensitive and specific surveillance of SARS-CoV-2 humoral immunity.
Stern D, Meyer TC, Treindl F, Mages HW, Krüger M, Skiba M, Krüger JP, Zobel CM, Schreiner M, Grossegesse M, Rinner T, Peine C, Stoliaroff-Pépin A, Harder T, Hofmann N, Michel J, Nitsche A, Stahlberg S, Kneuer A, Sandoni A, Kubisch U, Schlaud M, Mankertz A, Schwarz T, Corman VM, Müller MA, Drosten C, de la Rosa K, Schaade L, Dorner MB, Dorner BG. Stern D, et al. Sci Rep. 2023 Dec 9;13(1):21846. doi: 10.1038/s41598-023-48581-9. Sci Rep. 2023. PMID: 38071261 Free PMC article.
Kinetics of dried blood spot-measured anti-SARS-CoV2 Spike IgG in mRNA-vaccinated healthcare workers.
Puccini L, Fantini M, Biagetti C, Angelini R, Dirani G, Grumiro L, Schiavone P, Sparacino M, Semprini S, Sambri V, Cricca M. Puccini L, et al. Front Microbiol. 2023 Mar 1;14:1130677. doi: 10.3389/fmicb.2023.1130677. eCollection 2023. Front Microbiol. 2023. PMID: 36937271 Free PMC article.
Seropositivity and risk factors for SARS-CoV-2 infection in a South Asian community in Ontario: a cross-sectional analysis of a prospective cohort study.
Anand SS, Arnold C, Bangdiwala SI, Bolotin S, Bowdish D, Chanchlani R, de Souza RJ, Desai D, Kandasamy S, Khan F, Khan Z, Langlois MA, Limbachia J, Lear SA, Loeb M, Loh L, Manoharan B, Nakka K, Pelchat M, Punthakee Z, Schulze KM, Williams N, Wahi G. Anand SS, et al. CMAJ Open. 2022 Jul 5;10(3):E599-E609. doi: 10.9778/cmajo.20220031. Print 2022 Jul-Sep. CMAJ Open. 2022. PMID: 35790229 Free PMC article.
Cohort profile: investigating SARS-CoV-2 infection and the health and psychosocial impact of the COVID-19 pandemic in the Canadian CHILD Cohort.
Azeez R, Lotoski L, Dubeau A, Rodriguez N, Reyna ME, Freitas T, Goguen S, Medeleanu M, Winsor GL, Brinkman FSL, Cameron EE, Roos L, Simons E, Moraes TJ, Mandhane PJ, Turvey SE, Bolotin S, Wright K, McNeil D, Patrick DM, Bullard J, Langlois MA, Arnold CR, Galipeau Y, Pelchat M, Doucas N, Subbarao P, Azad MB. Azeez R, et al. Epidemiol Health. 2023;45:e2023091. doi: 10.4178/epih.e2023091. Epub 2023 Oct 13. Epidemiol Health. 2023. PMID: 37857338 Free PMC article.
Does school reopening affect SARS-CoV-2 seroprevalence among school-age children in Milan?
Barcellini L, Forlanini F, Sangiorgio A, Gambacorta G, Alberti L, Meta A, Gaia P, Amendola A, Tanzi E, Massa V, Borghi E, Fabiano V, Zuccotti GV. Barcellini L, et al. PLoS One. 2021 Sep 2;16(9):e0257046. doi: 10.1371/journal.pone.0257046. eCollection 2021. PLoS One. 2021. PMID: 34473776 Free PMC article.

See all "Cited by" articles

References

1. Wu F, Zhao S, Yu B, Chen Y-M, Wang W, Song Z-G, et al.. A new coronavirus associated with human respiratory disease in China. Nature. 2020;579(7798):265–9. doi: 10.1038/s41586-020-2008-3 - DOI - PMC - PubMed
1. Centre for Systems and Science Engineering at John Hopkins University. COVID-19 dashboard [cited 2021 January 19]. Available from: https://coronavirus.jhu.edu/map.html.
1. Corman VM, Landt O, Kaiser M, Molenkamp R, Meijer A, Chu DK, et al.. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro Surveillance. 2020;25(3):23–30. doi: 10.2807/1560-7917.ES.2020.25.3.2000045 - DOI - PMC - PubMed
1. Mallett S, Allen AJ, Graziadio S, Taylor SA, Sakai NS, Green K, et al.. At what times during infection is SARS-CoV-2 detectable and no longer detectable using RT-PCR-based tests? A systematic review of individual participant data. BMC Medicine. 2020;18(1):346. doi: 10.1186/s12916-020-01810-8 - DOI - PMC - PubMed
1. Becker MG, Taylor T, Kiazyk S, Cabiles DR, Meyers AFA, Sandstrom PA. Recommendations for sample pooling on the Cepheid GeneXpert(R) system using the Cepheid Xpert(R) Xpress SARS-CoV-2 assay. PLoS ONE. 2020;15(11):e0241959. doi: 10.1371/journal.pone.0241959 - DOI - PMC - PubMed

Publication types

Actions
Actions

MeSH terms

Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions

Substances

Actions
Actions

LinkOut - more resources

Full Text Sources
Medical
- MedlinePlus Consumer Health Information
- MedlinePlus Health Information
Miscellaneous
- NCI CPTAC Assay Portal

Save citation to file

Email citation

Add to Collections

Add to My Bibliography

Your saved search

Create a file for external citation management software

Your RSS Feed

Dried blood spot specimens for SARS-CoV-2 antibody testing: A multi-site, multi-assay comparison

Affiliations

Dried blood spot specimens for SARS-CoV-2 antibody testing: A multi-site, multi-assay comparison

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

Similar articles

Cited by

References

Publication types

MeSH terms

Substances

LinkOut - more resources

Full Text Sources

Medical

Miscellaneous