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Lücke, A.C.* ; Vom Hemdt, A.* ; Wieseler, J.* ; Fischer, C.* ; Feldmann, M.* ; Rothenfußer, S. ; Drexler, J.F.* ; Kümmerer, B.M.*

High-throughput platform for detection of neutralizing antibodies using flavivirus reporter replicon particles.

Viruses 14:346 (2022)
Verlagsversion Forschungsdaten DOI PMC
Open Access Gold
Creative Commons Lizenzvertrag
Flavivirus outbreaks require fast and reliable diagnostics that can be easily adapted to newly emerging and re-emerging flaviviruses. Due to the serological cross-reactivity among fla-vivirus antibodies, neutralization tests (NT) are considered the gold standard for sero-diagnostics. Here, we first established wild-type single-round infectious virus replicon particles (VRPs) by packaging a yellow fever virus (YFV) replicon expressing Gaussia luciferase (Gluc) with YFV structural proteins in trans using a double subgenomic Sindbis virus (SINV) replicon. The latter expressed the YFV envelope proteins prME via the first SINV subgenomic promoter and the capsid protein via a second subgenomic SINV promoter. VRPs were produced upon co-electroporation of replicon and packaging RNA. Introduction of single restriction enzyme sites in the packaging construct flanking the prME sequence easily allowed to exchange the prME moiety resulting in chimeric VRPs that have the surface proteins of other flaviviruses including dengue virus 1-4, Zika virus, West Nile virus, and tick-borne encephalitis virus. Besides comparing the YF-VRP based NT assay to a YF reporter virus NT assay, we analyzed the neutralization efficiencies of different human anti-fla-vivirus sera or a monoclonal antibody against all established VRPs. The assays were performed in a 96-well high-throughput format setting with Gluc as readout in comparison to classical plaque reduction NTs indicating that the VRP-based NT assays are suitable for high-throughput analyses of neutralizing flavivirus antibodies.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Korrespondenzautor
Schlagwörter Diagnostics ; Flavivirus ; High-throughput ; Neutralization Assay ; Virus Replicon Particles; West-nile-virus; Zika Virus; Rna; Infection; Dengue; Cdna; Construction; Elements; Culture; Vaccine
ISSN (print) / ISBN 1999-4915
e-ISSN 1999-4915
Zeitschrift Viruses
Quellenangaben Band: 14, Heft: 2, Seiten: , Artikelnummer: 346 Supplement: ,
Verlag MDPI
Verlagsort St Alban-anlage 66, Ch-4052 Basel, Switzerland
Nichtpatentliteratur Publikationen
Begutachtungsstatus Peer reviewed
Institut(e) Unit for Clinical Pharmacology (KKG-EKLiP)
Förderungen Deutsche Forschungsgemeinschaft (DFG, German Research foundation)
European Union's Horizon 2020 research and innovation program through the ZIKAlliance project