. 2020 Dec 17;59(1):e02204-20.

doi: 10.1128/JCM.02204-20. Print 2020 Dec 17.

COVID-19 Serology at Population Scale: SARS-CoV-2-Specific Antibody Responses in Saliva

Nora Pisanic^#¹, Pranay R Randad^#¹, Kate Kruczynski¹, Yukari C Manabe^{2

3}, David L Thomas², Andrew Pekosz^{1

4}, Sabra L Klein^{4

5

6}, Michael J Betenbaugh⁷, William A Clarke³, Oliver Laeyendecker^{2

8

9}, Patrizio P Caturegli^{3

10}, H Benjamin Larman^{3

10}, Barbara Detrick^{3

10}, Jessica K Fairley¹¹, Amy C Sherman¹², Nadine Rouphael¹², Srilatha Edupuganti¹², Douglas A Granger¹³, Steve W Granger¹⁴, Matthew H Collins¹², Christopher D Heaney^{15

6

8}

Affiliations

¹ Department of Environmental Health and Engineering, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, USA.
² Division of Infectious Diseases, Department of Medicine, School of Medicine, Johns Hopkins University, Baltimore, Maryland, USA.
³ Department of Pathology, School of Medicine, Johns Hopkins University, Baltimore, Maryland, USA.
⁴ Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, USA.
⁵ Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, USA.
⁶ Department of International Health, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, USA.
⁷ Department of Chemical and Biomolecular Engineering, Whiting School of Engineering, Johns Hopkins University, Baltimore, Maryland, USA.
⁸ Department of Epidemiology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, USA.
⁹ Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Baltimore, Maryland, USA.
¹⁰ Division of Immunology, Department of Pathology, School of Medicine, Johns Hopkins University, Baltimore, Maryland, USA.
¹¹ Department of Medicine, Emory University School of Medicine, Atlanta, Georgia, USA.
¹² The Hope Clinic of the Emory Vaccine Center, Division of Infectious Diseases, Department of Medicine, Emory University School of Medicine, Decatur, Georgia, USA.
¹³ Institute for Interdisciplinary Salivary Bioscience Research, University of California at Irvine, Irvine, California, USA.
¹⁴ Salimetrics, LLC, Carlsbad, California, USA.
¹⁵ Department of Environmental Health and Engineering, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, USA cheaney1@jhu.edu.

^# Contributed equally.

PMID: 33067270
PMCID: PMC7771435
DOI: 10.1128/JCM.02204-20

COVID-19 Serology at Population Scale: SARS-CoV-2-Specific Antibody Responses in Saliva

Nora Pisanic et al. J Clin Microbiol. 2020.

. 2020 Dec 17;59(1):e02204-20.

doi: 10.1128/JCM.02204-20. Print 2020 Dec 17.

Authors

Affiliations

¹ Department of Environmental Health and Engineering, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, USA.
² Division of Infectious Diseases, Department of Medicine, School of Medicine, Johns Hopkins University, Baltimore, Maryland, USA.
³ Department of Pathology, School of Medicine, Johns Hopkins University, Baltimore, Maryland, USA.
⁴ Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, USA.
⁵ Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, USA.
⁶ Department of International Health, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, USA.
⁷ Department of Chemical and Biomolecular Engineering, Whiting School of Engineering, Johns Hopkins University, Baltimore, Maryland, USA.
⁸ Department of Epidemiology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, USA.
⁹ Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Baltimore, Maryland, USA.
¹⁰ Division of Immunology, Department of Pathology, School of Medicine, Johns Hopkins University, Baltimore, Maryland, USA.
¹¹ Department of Medicine, Emory University School of Medicine, Atlanta, Georgia, USA.
¹² The Hope Clinic of the Emory Vaccine Center, Division of Infectious Diseases, Department of Medicine, Emory University School of Medicine, Decatur, Georgia, USA.
¹³ Institute for Interdisciplinary Salivary Bioscience Research, University of California at Irvine, Irvine, California, USA.
¹⁴ Salimetrics, LLC, Carlsbad, California, USA.
¹⁵ Department of Environmental Health and Engineering, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, USA cheaney1@jhu.edu.

^# Contributed equally.

PMID: 33067270
PMCID: PMC7771435
DOI: 10.1128/JCM.02204-20

Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of an ongoing pandemic that has infected over 36 million and killed over 1 million people. Informed implementation of government public health policies depends on accurate data on SARS-CoV-2 immunity at a population scale. We hypothesized that detection of SARS-CoV-2 salivary antibodies could serve as a noninvasive alternative to serological testing for monitoring of SARS-CoV-2 infection and seropositivity at a population scale. We developed a multiplex SARS-CoV-2 antibody immunoassay based on Luminex technology that comprised 12 CoV antigens, mostly derived from SARS-CoV-2 nucleocapsid (N) and spike (S). Saliva and sera collected from confirmed coronavirus disease 2019 (COVID-19) cases and from the pre-COVID-19 era were tested for IgG, IgA, and IgM to the antigen panel. Matched saliva and serum IgG responses (n = 28) were significantly correlated. The salivary anti-N IgG response resulted in the highest sensitivity (100%), exhibiting a positive response in 24/24 reverse transcription-PCR (RT-PCR)-confirmed COVID-19 cases sampled at >14 days post-symptom onset (DPSO), whereas the salivary anti-receptor binding domain (RBD) IgG response yielded 100% specificity. Temporal kinetics of IgG in saliva were consistent with those observed in blood and indicated that most individuals seroconvert at around 10 DPSO. Algorithms employing a combination of the IgG responses to N and S antigens result in high diagnostic accuracy (100%) by as early as 10 DPSO. These results support the use of saliva-based antibody testing as a noninvasive and scalable alternative to blood-based antibody testing.

Keywords: COVID-19; SARS-CoV-2; antibody test; diagnostics; immunoserology; multiplex; oral fluid; saliva; serology.

PubMed Disclaimer

Figures

**FIG 1**
Correlation between matched SARS-CoV-2-specific IgG responses in saliva and serum (n = 28). The Pearson correlation coefficient is provided for IgG responses to each antigen. Sino Biol., Sino Biological; NAC, Native Antigen Company; N, nucleocapsid protein; ECD, ectodomain; S1/S2, S1 or S2 subunit of the spike protein; RBD, receptor binding domain; (h), produced in human cells; (i), produced in insect cells; MFI, median fluorescence intensity.

**FIG 2**
Correlation between matched salivary and serum SARS-CoV-2-specific IgA responses (n = 26). The Pearson correlation coefficient is provided for IgA responses to each antigen. P values are provided for statistically significant correlations only (*P <* 0.05). Sino Biol., Sino Biological; NAC, Native Antigen Company; N, nucleocapsid protein; ECD, ectodomain; S1/S2, S1 or S2 subunit of the spike protein; RBD, receptor binding domain; (h), produced in human cells; (i), produced in insect cells; MFI, median fluorescence intensity.

**FIG 3**
Correlation between matched salivary and serum SARS-CoV-2-specific IgM responses (n = 26). The Pearson correlation coefficient is provided for IgM responses to each antigen. P values are provided for statistically significant correlations only (*P <* 0.05). Sino Biol., Sino Biological; NAC, Native Antigen Company; N, nucleocapsid protein; ECD, ectodomain; S1/S2, S1 or S2 subunit of the spike protein; RBD, receptor binding domain; (h), produced in human cells; (i), produced in insect cells; MFI, median fluorescence intensity.

**FIG 4**
Assay sensitivity and specificity for each SARS-CoV-2 antigen and antibody isotype using saliva (a) and serum (b). Samples collected from individuals with RT-PCR-confirmed prior SARS-CoV-2 infection are stratified by time since symptom onset. Darker shades of green indicate higher and darker shades of red indicate lower sensitivity and specificity. Sino Biol., Sino Biological; NAC, Native Antigen Company; N, nucleocapsid protein; ECD, ectodomain; S1/S2, S1 or S2 subunit of the spike protein; RBD, receptor binding domain; (h), produced in human cells; (i), produced in insect cells.

**FIG 5**
Comparison of saliva and serum SARS-CoV-2 antigen-specific IgG responses by days post-symptom onset (DPSO). The trajectories of IgG responses (red solid lines) and confidence intervals (semitransparent background) were estimated using a LOESS curve. Dashed red lines indicate cutoff values for each antigen. Sino Biol., Sino Biological; NAC, Native Antigen Company; N, nucleocapsid protein; ECD, ectodomain; S1/S2, S1 or S2 subunit of the spike protein; RBD, receptor binding domain; (h), produced in human cells; (i), produced in insect cells; MFI, median fluorescence intensity; a.u., arbitrary units.

See this image and copyright information in PMC

Update of

COVID-19 serology at population scale: SARS-CoV-2-specific antibody responses in saliva.
Randad PR, Pisanic N, Kruczynski K, Manabe YC, Thomas D, Pekosz A, Klein SL, Betenbaugh MJ, Clarke WA, Laeyendecker O, Caturegli PP, Larman HB, Detrick B, Fairley JK, Sherman AC, Rouphael N, Edupuganti S, Granger DA, Granger SW, Collins M, Heaney CD. Randad PR, et al. medRxiv [Preprint]. 2020 May 26:2020.05.24.20112300. doi: 10.1101/2020.05.24.20112300. medRxiv. 2020. Update in: J Clin Microbiol. 2020 Dec 17;59(1):e02204-20. doi: 10.1128/JCM.02204-20. PMID: 32511537 Free PMC article. Updated. Preprint.

References

1. Dong E, Du H, Gardner L. 2020. An interactive Web-based dashboard to track COVID-19 in real time. Lancet Infect Dis 20:533–534. doi:10.1016/S1473-3099(20)30120-1. - DOI - PMC - PubMed
1. Angulo FJ, Finelli L, Swerdlow DL. 2020. Reopening society and the need for real-time assessment of COVID-19 at the community level. JAMA 323:2247–2248. doi:10.1001/jama.2020.7872. - DOI - PubMed
1. Gronvall G, Connel N, Kobokovich A, West R, Warmbrod K, Shearer M, Mullen L, Inglesby T. 2020. Developing a national strategy for serology (antibody testing) in the United States. John Hopkins Bloomberg School of Public Health, Baltimore, MD.
1. Dyal JW, Grant MP, Broadwater K, Bjork A, Waltenburg MA, Gibbins JD, Hale C, Silver M, Fischer M, Steinberg J, Basler CA, Jacobs JR, Kennedy ED, Tomasi S, Trout D, Hornsby-Myers J, Oussayef NL, Delaney LJ, Patel K, Shetty V, Kline KE, Schroeder B, Herlihy RK, House J, Jervis R, Clayton JL, Ortbahn D, Austin C, Berl E, Moore Z, Buss BF, Stover D, Westergaard R, Pray I, DeBolt M, Person A, Gabel J, Kittle TS, Hendren P, Rhea C, Holsinger C, Dunn J, Turabelidze G, Ahmed FS, deFijter S, Pedati CS, Rattay K, Smith EE, Luna-Pinto C, Cooley LA, et al. . 2020. COVID-19 among workers in meat and poultry processing facilities—19 states, April 2020. MMWR Morb Mortal Wkly Rep 69:557–561. doi:10.15585/mmwr.mm6918e3. - DOI - PubMed
1. Lloyd-Sherlock P, Ebrahim S, Geffen L, McKee M. 2020. Bearing the brunt of covid-19: older people in low and middle income countries. BMJ 368:m1052. doi:10.1136/bmj.m1052. - DOI - PubMed

Publication types

Actions
Actions

MeSH terms

Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions

Substances

Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions

Grants and funding

LinkOut - more resources

Full Text Sources
Other Literature Sources
- The Lens - Patent Citations Database
Medical
- 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

COVID-19 Serology at Population Scale: SARS-CoV-2-Specific Antibody Responses in Saliva

Affiliations

COVID-19 Serology at Population Scale: SARS-CoV-2-Specific Antibody Responses in Saliva

Authors

Affiliations

Abstract

Figures

Update of

References

Publication types

MeSH terms

Substances

Grants and funding

LinkOut - more resources

Full Text Sources

Other Literature Sources

Medical

Miscellaneous