A Novel Method to Reduce ELISA Serial Dilution Assay Workload Applied to SARS-CoV-2 and Seasonal HCoVs.

David Pattinson,Yoshihiro Kawaoka,Hongyu Rao,Shiho Chiba,Kiyoko Iwatsuki-Horimoto,Lizheng Guan,Keiko Mitamura,Masao Hagihara,Tomoyuki Uchida,Seiya Yamayoshi,Robert Presler,Nobuhiro Ikeda,Peter Jester,Peter Halfmann,Gabriele Neumann

doi:10.3390/v14030562

David Pattinson, Yoshihiro Kawaoka + Show 13 more

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https://doi.org/10.3390/v14030562

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Assays using ELISA measurements on serially diluted serum samples have been heavily used to measure serum reactivity to SARS-CoV-2 antigens and are widely used in virology and elsewhere in biology. We test a method using Bayesian hierarchical modelling to reduce the workload of these assays and measure reactivity of SARS-CoV-2 and HCoV antigens to human serum samples collected before and during the COVID-19 pandemic. Inflection titers for SARS-CoV-2 full-length spike protein (S1S2), spike protein receptor-binding domain (RBD), and nucleoprotein (N) inferred from 3 spread-out dilutions correlated with those inferred from 8 consecutive dilutions with an R2 value of 0.97 or higher. We confirm existing findings showing a small proportion of pre-pandemic human serum samples contain cross-reactive antibodies to SARS-CoV-2 S1S2 and N, and that SARS-CoV-2 infection increases serum reactivity to the beta-HCoVs OC43 and HKU1 S1S2. In serial dilution assays, large savings in resources and/or increases in throughput can be achieved by reducing the number of dilutions measured and using Bayesian hierarchical modelling to infer inflection or endpoint titers. We have released software for conducting these types of analysis.

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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in December 2019 [1] and is the causative agent of coronavirus disease 2019 (COVID-19)
We tested human serum samples collected before and during the SARS-CoV-2 pandemic for antibodies to SARS-CoV-2 and HCoVs, and used this well-characterized dataset to test a computational approach for substantially reducing the number of serial dilutions needed to robustly measure inflection points of titration series
We measured the reactivity of human serum samples collected before the pandemic and from individuals who were PCR-positive for SARS-CoV-2 against SARS-CoV-2 S1S2, receptor-binding domain (RBD) and N antigens at a single serum dilution of 1:40 (Figure 1A)

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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in December 2019 [1] and is the causative agent of coronavirus disease 2019 (COVID-19). On 11 March 2020, the outbreak was declared a pandemic by the World Health Organization (WHO) [2] and at the time of writing there have been over 206 million confirmed SARS-CoV-2 infections and 4.3 million deaths attributed to the virus globally [3]. Four CoVs circulate continuously in humans (HCoVs), are distributed globally, and generally cause common cold or influenzalike illness [5]. These are the alpha-CoVs HCoV-229E (229E) and HCoV-NL63 (NL63), and the beta-CoVs HCoV-HKU1 (HKU1) and HCoV-OC43 (OC43). Two other beta-CoVs have infected humans: severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East Respiratory Syndrome Coronavirus (MERS-CoV) [6,7]. The SARS-CoV outbreak in 2003 was largely contained and MERS-CoV is not well adapted for ongoing human to human transmission

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