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A method to reduce ELISA serial dilution assay workload applied to SARS-CoV-2 and seasonal HCoVs

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

Posted 21 Sep 2021
medRxiv DOI: 10.1101/2021.09.13.21263523

Objectives 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 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. Methods We apply Bayesian hierarchical modelling to ELISA measurements of human serum samples against SARS-CoV-2 and HCoV antigens. Results Inflection titers for SARS-CoV-2 full-length spike protein (S1S2), spike protein receptor-binding domain (RBD), and nucleoprotein (N) inferred from three spread-out dilutions correlated with those inferred from eight 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. Conclusions 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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