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FluoRNT: A robust, efficient assay for the detection of neutralising antibodies against yellow fever virus 17D.
Scheck, Magdalena K; Lehmann, Lisa; Zaucha, Magdalena; Schwarzlmueller, Paul; Huber, Kristina; Pritsch, Michael; Barba-Spaeth, Giovanna; Thorn-Seshold, Oliver; Krug, Anne B; Endres, Stefan; Rothenfusser, Simon; Thorn-Seshold, Julia.
  • Scheck MK; Division of Clinical Pharmacology, University Hospital, LMU Munich, Munich, Germany.
  • Lehmann L; Division of Clinical Pharmacology, University Hospital, LMU Munich, Munich, Germany.
  • Zaucha M; Division of Clinical Pharmacology, University Hospital, LMU Munich, Munich, Germany.
  • Schwarzlmueller P; Division of Clinical Pharmacology, University Hospital, LMU Munich, Munich, Germany.
  • Huber K; Division of Infectious Diseases and Tropical Medicine, University Hospital, LMU Munich, Munich, Germany.
  • Pritsch M; Division of Infectious Diseases and Tropical Medicine, University Hospital, LMU Munich, Munich, Germany.
  • Barba-Spaeth G; German Center for Infection Research (DZIF), Partner Site Munich, Munich, Germany.
  • Thorn-Seshold O; Structural Virology Unit and CNRS UMR 3569, Virology Department, Institute Pasteur, Paris, France.
  • Krug AB; Faculty of Chemistry and Pharmacy, LMU Munich, Munich, Germany.
  • Endres S; Institute for Immunology, Biomedical Center, Faculty of Medicine, LMU Munich, Planegg-Martinsried, Germany.
  • Rothenfusser S; Division of Clinical Pharmacology, University Hospital, LMU Munich, Munich, Germany.
  • Thorn-Seshold J; Unit Clinical Pharmacology (EKliP), Helmholtz Center for Environmental Health, Munich, Germany.
PLoS One ; 17(2): e0262149, 2022.
Article in English | MEDLINE | ID: covidwho-1910485
ABSTRACT
There is an urgent need for better diagnostic and analytical methods for vaccine research and infection control in virology. This has been highlighted by recently emerging viral epidemics and pandemics (Zika, SARS-CoV-2), and recurring viral outbreaks like the yellow fever outbreaks in Angola and the Democratic Republic of Congo (2016) and in Brazil (2016-2018). Current assays to determine neutralising activity against viral infections in sera are costly in time and equipment and suffer from high variability. Therefore, both basic infection research and diagnostic population screenings would benefit from improved methods to determine virus-neutralising activity in patient samples. Here we describe a robust, objective, and scalable Fluorescence Reduction Neutralisation Test (FluoRNT) for yellow fever virus, relying on flow cytometric detection of cells infected with a fluorescent Venus reporter containing variant of the yellow fever vaccine strain 17D (YF-17D-Venus). It accurately measures neutralising antibody titres in human serum samples within as little as 24 h. Samples from 32 vaccinees immunised with YF-17D were tested for neutralising activity by both a conventional focus reduction neutralisation test (FRNT) and FluoRNT. Both types of tests proved to be equally reliable for the detection of neutralising activity, however, FluoRNT is significantly more precise and reproducible with a greater dynamic range than conventional FRNT. The FluoRNT assay protocol is substantially faster, easier to control, and cheaper in per-assay costs. FluoRNT additionally reduces handling time minimising exposure of personnel to patient samples. FluoRNT thus brings a range of desirable features that can accelerate and standardise the measurement of neutralising anti-yellow fever virus antibodies. It could be used in applications ranging from vaccine testing to large cohort studies in systems virology and vaccinology. We also anticipate the potential to translate the methodology and analysis of FluoRNT to other flaviviruses such as West Nile, Dengue and Zika or to RNA viruses more generally.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Yellow Fever / Yellow fever virus / Antibodies, Neutralizing Type of study: Cohort study / Diagnostic study / Observational study / Prognostic study Topics: Vaccines / Variants Limits: Animals / Humans Language: English Journal: PLoS One Journal subject: Science / Medicine Year: 2022 Document Type: Article Affiliation country: Journal.pone.0262149

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Yellow Fever / Yellow fever virus / Antibodies, Neutralizing Type of study: Cohort study / Diagnostic study / Observational study / Prognostic study Topics: Vaccines / Variants Limits: Animals / Humans Language: English Journal: PLoS One Journal subject: Science / Medicine Year: 2022 Document Type: Article Affiliation country: Journal.pone.0262149