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A Novel Method to Reduce ELISA Serial Dilution Assay Workload Applied to SARS-CoV-2 and Seasonal HCoVs.
Pattinson, David; Jester, Peter; Guan, Lizheng; Yamayoshi, Seiya; Chiba, Shiho; Presler, Robert; Rao, Hongyu; Iwatsuki-Horimoto, Kiyoko; Ikeda, Nobuhiro; Hagihara, Masao; Uchida, Tomoyuki; Mitamura, Keiko; Halfmann, Peter; Neumann, Gabriele; Kawaoka, Yoshihiro.
  • Pattinson D; Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI 53706, USA.
  • Jester P; Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI 53706, USA.
  • Guan L; Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI 53706, USA.
  • Yamayoshi S; Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo 108-0071, Japan.
  • Chiba S; The Research Center for Global Viral Diseases, Research Institute, National Center for Global Health and Medicine, Tokyo 162-8665, Japan.
  • Presler R; Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI 53706, USA.
  • Rao H; Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI 53706, USA.
  • Iwatsuki-Horimoto K; Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI 53706, USA.
  • Ikeda N; Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo 108-0071, Japan.
  • Hagihara M; Department of General Internal Medicine, Eiju General Hospital, Tokyo 104-0045, Japan.
  • Uchida T; Department of Hematology, Eiju General Hospital, Tokyo 104-0045, Japan.
  • Mitamura K; Department of Hematology, Eiju General Hospital, Tokyo 104-0045, Japan.
  • Halfmann P; Division of Infection Control, Eiju General Hospital, Tokyo 104-0045, Japan.
  • Neumann G; Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI 53706, USA.
  • Kawaoka Y; Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI 53706, USA.
Viruses ; 14(3)2022 03 09.
Article in English | MEDLINE | ID: covidwho-1732253
ABSTRACT
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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Full text: Available Collection: International databases Database: MEDLINE Main subject: SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study / Randomized controlled trials Limits: Humans Language: English Year: 2022 Document Type: Article Affiliation country: V14030562

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Full text: Available Collection: International databases Database: MEDLINE Main subject: SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study / Randomized controlled trials Limits: Humans Language: English Year: 2022 Document Type: Article Affiliation country: V14030562