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Open AccessArticle

Improved Detection of Antibodies against SARS-CoV-2 by Microsphere-Based Antibody Assay

1
State Key Laboratory for Emerging Infectious Diseases, Carol Yu Centre for Infection, Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, China
2
Department of Microbiology, Queen Mary Hospital, Hong Kong, China
3
Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, China
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(18), 6595; https://doi.org/10.3390/ijms21186595
Received: 14 August 2020 / Revised: 1 September 2020 / Accepted: 3 September 2020 / Published: 9 September 2020
(This article belongs to the Special Issue Molecular Research in Emerging Viruses 2020)
Currently available COVID-19 antibody tests using enzyme immunoassay (EIA) or immunochromatographic assay have variable sensitivity and specificity. Here, we developed and evaluated a novel microsphere-based antibody assay (MBA) for detecting immunoglobulin G (IgG) against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleoprotein (NP) and spike protein receptor binding domain (RBD). The seropositive cutoff value was set using a cohort of 294 anonymous serum specimens collected in 2018. The specificity was assessed using serum specimens collected from organ donors or influenza patients before 2020. Seropositive rate was determined among COVID-19 patients. Time-to-seropositivity and signal-to-cutoff (S/CO) ratio were compared between MBA and EIA. MBA had a specificity of 100% (93/93; 95% confidence interval (CI), 96–100%) for anti-NP IgG, 98.9% (92/93; 95% CI 94.2–100%) for anti-RBD IgG. The MBA seropositive rate for convalescent COVID-19 patients was 89.8% (35/39) for anti-NP IgG and 79.5% (31/39) for anti-RBD IgG. The time-to-seropositivity was shorter with MBA than EIA. MBA could better differentiate between COVID-19 patients and negative controls with higher S/CO ratio for COVID-19 patients, lower S/CO ratio with negative controls and fewer specimens in the equivocal range. MBA is robust, simple and is suitable for clinical microbiology laboratory for the accurate determination of anti-SARS-CoV-2 antibodies for diagnosis, serosurveillance, and vaccine trials. View Full-Text
Keywords: COVID-19; SARS-CoV-2; serology; flow cytometry; antibody assay COVID-19; SARS-CoV-2; serology; flow cytometry; antibody assay
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MDPI and ACS Style

Fong, C.H.-Y.; Cai, J.-P.; Dissanayake, T.K.; Chen, L.-L.; Choi, C.Y.-K.; Wong, L.-H.; Ng, A.C.-K.; Pang, P.K.P.; Ho, D.T.-Y.; Poon, R.W.-S.; Chung, T.W.-H.; Sridhar, S.; Chan, K.-H.; Chan, J.F.-W.; Hung, I.F.-N.; Yuen, K.-Y.; To, K.K.-W. Improved Detection of Antibodies against SARS-CoV-2 by Microsphere-Based Antibody Assay. Int. J. Mol. Sci. 2020, 21, 6595. https://doi.org/10.3390/ijms21186595

AMA Style

Fong CH-Y, Cai J-P, Dissanayake TK, Chen L-L, Choi CY-K, Wong L-H, Ng AC-K, Pang PKP, Ho DT-Y, Poon RW-S, Chung TW-H, Sridhar S, Chan K-H, Chan JF-W, Hung IF-N, Yuen K-Y, To KK-W. Improved Detection of Antibodies against SARS-CoV-2 by Microsphere-Based Antibody Assay. International Journal of Molecular Sciences. 2020; 21(18):6595. https://doi.org/10.3390/ijms21186595

Chicago/Turabian Style

Fong, Carol H.-Y.; Cai, Jian-Piao; Dissanayake, Thrimendra K.; Chen, Lin-Lei; Choi, Charlotte Y.-K.; Wong, Lok-Hin; Ng, Anthony C.-K.; Pang, Polly K.P.; Ho, Deborah T.-Y.; Poon, Rosana W.-S.; Chung, Tom W.-H.; Sridhar, Siddharth; Chan, Kwok-Hung; Chan, Jasper F.-W.; Hung, Ivan F.-N.; Yuen, Kwok-Yung; To, Kelvin K.-W. 2020. "Improved Detection of Antibodies against SARS-CoV-2 by Microsphere-Based Antibody Assay" Int. J. Mol. Sci. 21, no. 18: 6595. https://doi.org/10.3390/ijms21186595

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