Serological assays for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), March 2020

Ranawaka Apm Perera, Chris Kp Mok, Owen Ty Tsang, Huibin Lv, Ronald Lw Ko, Nicholas C Wu, Meng Yuan, Wai Shing Leung, Jacky Mc Chan, Thomas Sh Chik, Chris Yc Choi, Kathy Leung, Kin Ho Chan, Karl Ck Chan, Ka-Chi Li, Joseph T Wu, Ian A Wilson, Arnold S Monto, Leo Lm Poon, Malik Peiris, Ranawaka Apm Perera, Chris Kp Mok, Owen Ty Tsang, Huibin Lv, Ronald Lw Ko, Nicholas C Wu, Meng Yuan, Wai Shing Leung, Jacky Mc Chan, Thomas Sh Chik, Chris Yc Choi, Kathy Leung, Kin Ho Chan, Karl Ck Chan, Ka-Chi Li, Joseph T Wu, Ian A Wilson, Arnold S Monto, Leo Lm Poon, Malik Peiris

Abstract

BackgroundThe ongoing coronavirus disease (COVID-19) pandemic has major impacts on health systems, the economy and society. Assessing infection attack rates in the population is critical for estimating disease severity and herd immunity which is needed to calibrate public health interventions. We have previously shown that it is possible to achieve this in real time to impact public health decision making.AimOur objective was to develop and evaluate serological assays applicable in large-scale sero-epidemiological studies.MethodsWe developed an ELISA to detect IgG and IgM antibodies to the receptor-binding domain (RBD) of the spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We evaluated its sensitivity and specificity in combination with confirmatory microneutralisation (MN) and 90% plaque reduction neutralisation tests (PRNT90) in 51 sera from 24 patients with virologically confirmed COVID-19 and in age-stratified sera from 200 healthy controls.ResultsIgG and IgM RBD ELISA, MN and PRNT90 were reliably positive after 29 days from illness onset with no detectable cross-reactivity in age-stratified controls. We found that PRNT90 tests were more sensitive in detecting antibody than MN tests carried out with the conventional 100 tissue culture infectious dose challenge. Heparinised plasma appeared to reduce the infectivity of the virus challenge dose and may confound interpretation of neutralisation test.ConclusionUsing IgG ELISA based on the RBD of the spike protein to screen sera for SARS-CoV-2 antibody, followed by confirmation using PRNT90, is a valid approach for large-scale sero-epidemiology studies.

Keywords: COVD19; ELISA; SARS-CoV-2; neutralization; receptor binding domain; serology.

Conflict of interest statement

Conflict of interest: None declared.

Figures

Figure 1
Figure 1
Antibody responses of the COVID-19 patient cohort in relation to duration of illness, Hong Kong, March 2020 (n = 24 patients, 51 sera)
Figure 2
Figure 2
Kinetics of antibody response in individual patients with SARS-CoV-2 infection by days after illness onset, Hong Kong, March 2020 (n = 17 patients)
Figure 3
Figure 3
Correlation between the ELISA test results and microneutralisation and plaque reduction neutralisation results in COVID-19 patients, Hong Kong, March 2020 (n = 24 patients, 51 sera)
Figure 4
Figure 4
Correlation between microneutralisation and IgG ELISA in pairs of plasma and serum specimens collected on the same day from COVID-19 patients, Hong Kong, March 2020 (n = 16 pairs of serum and plasma)

References

    1. Zhu N, Zhang D, Wang W, Li X, Yang B, Song J, et al. A novel coronavirus from patients with pneumonia in China, 2019. N Engl J Med. 2020;382(8):727-33. 10.1056/NEJMoa2001017
    1. Word Health Organization (WHO). Coronavirus disease 2019 (COVID-19): Situation report 92. Geneva: WHO. [Accessed: 22 Apr 2020]. Available from:
    1. Battegay M, Kuehl R, Tschudin-Sutter S, Hirsch HH, Widmer AF, Neher RA. 2019-novel Coronavirus (2019-nCoV): estimating the case fatality rate - a word of caution. Swiss Med Wkly. 2020;150:w20203. 10.4414/smw.2020.20203
    1. Wu JT, Ma ES, Lee CK, Chu DK, Ho PL, Shen AL, et al. The infection attack rate and severity of 2009 pandemic H1N1 influenza in Hong Kong. Clin Infect Dis. 2010;51(10):1184-91. 10.1086/656740
    1. Wong JY, Kelly H, Ip DK, Wu JT, Leung GM, Cowling BJ. Case fatality risk of influenza A (H1N1pdm09): a systematic review. Epidemiology. 2013;24(6):830-41. 10.1097/EDE.0b013e3182a67448
    1. Monto AS, Malosh RE, Petrie JG, Thompson MG, Ohmit SE. Frequency of acute respiratory illnesses and circulation of respiratory viruses in households with children over 3 surveillance seasons. J Infect Dis. 2014;210(11):1792-9. 10.1093/infdis/jiu327
    1. Ekiert DC, Friesen RH, Bhabha G, Kwaks T, Jongeneelen M, Yu W, et al. A highly conserved neutralizing epitope on group 2 influenza A viruses. Science. 2011;333(6044):843-50. 10.1126/science.1204839
    1. Yuan M, Wu NC, Zhu X, Lee CD, So RTY, Lv H, et al. A highly conserved cryptic epitope in the receptor-binding domains of SARS-CoV-2 and SARS-CoV. Science. 2020;eabb7269. 10.1126/science.abb7269
    1. Moritz CP, Tholance Y, Lassablière F, Camdessanché JP, Antoine JC. Reducing the risk of misdiagnosis of indirect ELISA by normalizing serum-specific background noise: The example of detecting anti-FGFR3 autoantibodies. J Immunol Methods. 2019;466:52-6. 10.1016/j.jim.2019.01.004
    1. Reed LJ, Muench H. A simple method of estimating fifty percent end points. Am J Hyg. 1938;27:493-7.
    1. Choe PG, Perera RAPM, Park WB, Song KH, Bang JH, Kim ES, et al. MERS-CoV antibody responses 1 year after symptom onset, South Korea, 2015. Emerg Infect Dis. 2017;23(7):1079-84. 10.3201/eid2307.170310
    1. Chan CM, Tse H, Wong SS, Woo PC, Lau SK, Chen L, et al. Examination of seroprevalence of coronavirus HKU1 infection with S protein-based ELISA and neutralization assay against viral spike pseudotyped virus. J Clin Virol. 2009;45(1):54-60. 10.1016/j.jcv.2009.02.011
    1. Gorse GJ, Patel GB, Vitale JN, O’Connor TZ. Prevalence of antibodies to four human coronaviruses is lower in nasal secretions than in serum. Clin Vaccine Immunol. 2010;17(12):1875-80. 10.1128/CVI.00278-10
    1. Okba NMA, Müller MA, Li W, Wang C, GeurtsvanKessel CH, Corman VM, et al. Severe acute respiratory syndrome coronavirus 2-specific antibody responses in coronavirus disease 2019 patients. Emerg Infect Dis. 2020;26(7): Epub ahead of print. 10.3201/eid2607.200841
    1. Zhang B, Zhou X, Zhu C, Feng F, Qiu Y, Feng J, et al. Immune phenotyping based on neutrophil-to-lymphocyte ratio and IgG predicts disease severity and outcome for patients with COVID-19. medRxiv 2020. 10.1101/2020.03.12.20035048
    1. Wu JT, Leung K, Perera RA, Chu DK, Lee CK, Hung IF, et al. Inferring influenza infection attack rate from seroprevalence data. PLoS Pathog. 2014;10(4):e1004054. 10.1371/journal.ppat.1004054

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