Development and testing of a method for validating chemical inactivation of ebola virus

Kendra J. Alfson, Anthony Griffiths

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Complete inactivation of infectious Ebola virus (EBOV) is required before a sample may be removed from a Biosafety Level 4 laboratory. The United States Federal Select Agent Program regulations require that procedures used to demonstrate chemical inactivation must be validated in-house to confirm complete inactivation. The objective of this study was to develop a method for validating chemical inactivation of EBOV and then demonstrate the effectiveness of several commonly-used inactivation methods. Samples containing infectious EBOV (Zaire ebolavirus) in different matrices were treated, and the sample was diluted to limit the cytopathic effect of the inactivant. The presence of infectious virus was determined by assessing the cytopathic effect in Vero E6 cells. Crucially, this method did not result in a loss of infectivity in control samples, and we were able to detect less than five infectious units of EBOV (Zaire ebolavirus). We found that TRIzol LS reagent and RNA-Bee inactivated EBOV in serum; TRIzol LS reagent inactivated EBOV in clarified cell culture media; TRIzol reagent inactivated EBOV in tissue and infected Vero E6 cells; 10% neutral buffered formalin inactivated EBOV in tissue; and osmium tetroxide vapors inactivated EBOV on transmission electron microscopy grids. The methods described herein are easily performed and can be adapted to validate inactivation of viruses in various matrices and by various chemical methods.

Original languageEnglish (US)
Article number126
JournalViruses
Volume10
Issue number3
DOIs
StatePublished - Mar 13 2018
Externally publishedYes

Keywords

  • Chemical inactivation
  • Cytopathic effect
  • Ebola virus
  • Limit of detection
  • Viability testing

ASJC Scopus subject areas

  • Infectious Diseases
  • Virology

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