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Research Project: Viral Ecology of Henipaviruses in Endemic Settings and Intervention Strategies to Prevent their Spread to Domestic Animals

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Title: Inactivation of Ebola, Nipah, and Lassa viruses in tissue using neutral buffered formalin, MagMAXTM lysis/binding solution, or TriPure isolation reagent

Author
item DAVIES, KATHERINE - Oak Ridge Institute For Science And Education (ORISE)
item WELCH, STEPHEN - Centers For Disease Control And Prevention (CDC) - United States
item HARCOURT, BRIAN - Centers For Disease Control And Prevention (CDC) - United States
item SPIROPOULOU, CHRISTINA - Centers For Disease Control And Prevention (CDC) - United States
item SPENGLER, JESSICA - Centers For Disease Control And Prevention (CDC) - United States

Submitted to: Scientific Reports
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/22/2025
Publication Date: 12/31/2025
Citation: Davies, K.A., Welch, S.R., Harcourt, B.H., Spiropoulou, C.F., Spengler, J.R. 2025. Inactivation of Ebola, Nipah, and Lassa viruses in tissue using neutral buffered formalin, MagMAXTM lysis/binding solution, or TriPure isolation reagent. Scientific Reports. 16. Article 3712. https://doi.org/10.1038/s41598-025-33750-9.
DOI: https://doi.org/10.1038/s41598-025-33750-9

Interpretive Summary: There is a critical need to validate inactivation methods for tissues infected with high consequence pathogens such as Ebola, Lassa and Nipah viruses, to allow safe transfer from high-containment to lower containment for downstream analysis. Using tissues from experimentally infected animals, we evaluate viral load reduction after treatment with neutral buffered formalin, MagMax Lysis/Binding solution concentrate and TriPure isolation reagent. By using verified purification strategies, the cytotoxicity of these reagents was reduced to enable sensitive detection of any remaining infectious virus. We demonstrated >4 log reductions under the tested conditions, providing data for commonly used laboratory inactivation procedures to support safe handling of specimens in both research and outbreak settings.

Technical Abstract: Samples known or suspected to be infected with high-consequence viruses such as Ebola, Nipah, and Lassa must be handled under high biocontainment. Studies involving animal infections with these pathogens can generate tissues that require downstream analyses, including molecular assays and histopathology, which are more readily performed, or in some cases only feasible, at lower containment levels. Before removal from high containment for analyses at lower containment levels, specimens must undergo validated inactivation procedures. Here, we quantified viral load reduction in tissues infected with these pathogens following treatment with neutral-buffered formalin for 10 minutes, 1 hour, 3 days, or 7 days, and MagMAX lysis/binding solution concentrate or TriPure isolation reagent for 1- or 10-minutes. To ensure accurate detection of any residual infectious virus, samples were purified through resins or centrifugal filters to reduce reagent cytotoxicity and maximize volume of testable material. We demonstrated effective inactivation (=4 log10 reduction) of all three pathogens and quantified log-reduction values over multiple timepoints. These findings provide validation data to support safe handling of infectious tissues for research, field studies, and outbreak response.