Validated Methods for Inactivation of Tick-Borne Encephalitis Virus Compatible with Immune-Based and Enzymatic Downstream Analyses

Tick-Borne Encephalitis Virus (TBEV) is impacting public health in the Eurasian region, with increasing case numbers. There is, therefore, a need to expand research efforts and the corresponding infrastructure capacity. Since TBEV is classified as a risk group 3 organism in Switzerland, handling inf...

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Main Authors: Simone Leoni, Stephen L. Leib, Katharina Summermatter, Denis Grandgirard
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Viruses
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Online Access:https://www.mdpi.com/1999-4915/17/6/810
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author Simone Leoni
Stephen L. Leib
Katharina Summermatter
Denis Grandgirard
author_facet Simone Leoni
Stephen L. Leib
Katharina Summermatter
Denis Grandgirard
author_sort Simone Leoni
collection DOAJ
description Tick-Borne Encephalitis Virus (TBEV) is impacting public health in the Eurasian region, with increasing case numbers. There is, therefore, a need to expand research efforts and the corresponding infrastructure capacity. Since TBEV is classified as a risk group 3 organism in Switzerland, handling infectious material containing the virus is restricted to biosafety level 3 laboratories. In some instances, downstream analyses may need to be performed outside of the containment facility. It is, therefore, essential to validate effective inactivation protocols compatible with the safe and accurate processing of samples. This study evaluated UV irradiation, chemical treatment with detergents, and mechanical filtration as candidate methods to inactivate TBEV infectious samples, including culture supernatants and tissue homogenates, while preserving their compatibility for different assays. Among the methods tested, 45 s of UV irradiation or Triton-X100 at concentrations between 0.05% and 0.1% effectively inactivated TBEV while mostly preserving the integrity of the processed samples for immuno- or enzymatic assays. These findings establish safe and reliable procedures for advancing TBEV research beyond high-containment settings.
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spelling doaj-art-a45246d7685d4645ab60cc668f6b3fde2025-08-20T02:21:54ZengMDPI AGViruses1999-49152025-06-0117681010.3390/v17060810Validated Methods for Inactivation of Tick-Borne Encephalitis Virus Compatible with Immune-Based and Enzymatic Downstream AnalysesSimone Leoni0Stephen L. Leib1Katharina Summermatter2Denis Grandgirard3Institute for Infectious Diseases, University of Bern, 3001 Bern, SwitzerlandInstitute for Infectious Diseases, University of Bern, 3001 Bern, SwitzerlandInstitute for Infectious Diseases, University of Bern, 3001 Bern, SwitzerlandInstitute for Infectious Diseases, University of Bern, 3001 Bern, SwitzerlandTick-Borne Encephalitis Virus (TBEV) is impacting public health in the Eurasian region, with increasing case numbers. There is, therefore, a need to expand research efforts and the corresponding infrastructure capacity. Since TBEV is classified as a risk group 3 organism in Switzerland, handling infectious material containing the virus is restricted to biosafety level 3 laboratories. In some instances, downstream analyses may need to be performed outside of the containment facility. It is, therefore, essential to validate effective inactivation protocols compatible with the safe and accurate processing of samples. This study evaluated UV irradiation, chemical treatment with detergents, and mechanical filtration as candidate methods to inactivate TBEV infectious samples, including culture supernatants and tissue homogenates, while preserving their compatibility for different assays. Among the methods tested, 45 s of UV irradiation or Triton-X100 at concentrations between 0.05% and 0.1% effectively inactivated TBEV while mostly preserving the integrity of the processed samples for immuno- or enzymatic assays. These findings establish safe and reliable procedures for advancing TBEV research beyond high-containment settings.https://www.mdpi.com/1999-4915/17/6/810Tick-Borne Encephalitis Virus (TBEV)flavivirusinactivationimmuno-assayenzymatic assaysbiosafety
spellingShingle Simone Leoni
Stephen L. Leib
Katharina Summermatter
Denis Grandgirard
Validated Methods for Inactivation of Tick-Borne Encephalitis Virus Compatible with Immune-Based and Enzymatic Downstream Analyses
Viruses
Tick-Borne Encephalitis Virus (TBEV)
flavivirus
inactivation
immuno-assay
enzymatic assays
biosafety
title Validated Methods for Inactivation of Tick-Borne Encephalitis Virus Compatible with Immune-Based and Enzymatic Downstream Analyses
title_full Validated Methods for Inactivation of Tick-Borne Encephalitis Virus Compatible with Immune-Based and Enzymatic Downstream Analyses
title_fullStr Validated Methods for Inactivation of Tick-Borne Encephalitis Virus Compatible with Immune-Based and Enzymatic Downstream Analyses
title_full_unstemmed Validated Methods for Inactivation of Tick-Borne Encephalitis Virus Compatible with Immune-Based and Enzymatic Downstream Analyses
title_short Validated Methods for Inactivation of Tick-Borne Encephalitis Virus Compatible with Immune-Based and Enzymatic Downstream Analyses
title_sort validated methods for inactivation of tick borne encephalitis virus compatible with immune based and enzymatic downstream analyses
topic Tick-Borne Encephalitis Virus (TBEV)
flavivirus
inactivation
immuno-assay
enzymatic assays
biosafety
url https://www.mdpi.com/1999-4915/17/6/810
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AT stephenlleib validatedmethodsforinactivationoftickborneencephalitisviruscompatiblewithimmunebasedandenzymaticdownstreamanalyses
AT katharinasummermatter validatedmethodsforinactivationoftickborneencephalitisviruscompatiblewithimmunebasedandenzymaticdownstreamanalyses
AT denisgrandgirard validatedmethodsforinactivationoftickborneencephalitisviruscompatiblewithimmunebasedandenzymaticdownstreamanalyses