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FluoRNT: A robust, efficient assay for the detection of neutralising antibodies against yellow fever virus 17D
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 th...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8827462/ https://www.ncbi.nlm.nih.gov/pubmed/35139078 http://dx.doi.org/10.1371/journal.pone.0262149 |
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author | 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 |
author_facet | 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 |
author_sort | Scheck, Magdalena K. |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-8827462 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-88274622022-02-10 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 PLoS One Research Article 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. Public Library of Science 2022-02-09 /pmc/articles/PMC8827462/ /pubmed/35139078 http://dx.doi.org/10.1371/journal.pone.0262149 Text en © 2022 Scheck et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article 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 FluoRNT: A robust, efficient assay for the detection of neutralising antibodies against yellow fever virus 17D |
title | FluoRNT: A robust, efficient assay for the detection of neutralising antibodies against yellow fever virus 17D |
title_full | FluoRNT: A robust, efficient assay for the detection of neutralising antibodies against yellow fever virus 17D |
title_fullStr | FluoRNT: A robust, efficient assay for the detection of neutralising antibodies against yellow fever virus 17D |
title_full_unstemmed | FluoRNT: A robust, efficient assay for the detection of neutralising antibodies against yellow fever virus 17D |
title_short | FluoRNT: A robust, efficient assay for the detection of neutralising antibodies against yellow fever virus 17D |
title_sort | fluornt: a robust, efficient assay for the detection of neutralising antibodies against yellow fever virus 17d |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8827462/ https://www.ncbi.nlm.nih.gov/pubmed/35139078 http://dx.doi.org/10.1371/journal.pone.0262149 |
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