Abstract
The handling of highly pathogenic viruses, whether for diagnostic or research purposes, often requires an inactivation step. This article reviews available inactivation techniques published in peer-reviewed journals and their benefits and limitations in relation to the intended application. The bulk of highly pathogenic viruses are represented by enveloped RNA viruses belonging to the Togaviridae, Flaviviridae, Filoviridae, Arenaviridae, Hantaviridae, Peribunyaviridae, Phenuiviridae, Nairoviridae and Orthomyxoviridae families. Here, we summarize inactivation methods for these virus families that allow for subsequent molecular and serological analysis or vaccine development. The techniques identified here include: treatment with guanidium-based chaotropic salts, heat inactivation, photoactive compounds such as psoralens or 1.5-iodonaphtyl azide, detergents, fixing with aldehydes, UV-radiation, gamma irradiation, aromatic disulfides, beta-propiolacton and hydrogen peroxide. The combination of simple techniques such as heat or UV-radiation and detergents such as Tween-20, Triton X-100 or Sodium dodecyl sulfate are often sufficient for virus inactivation, but the efficiency may be affected by influencing factors including quantity of infectious particles, matrix constitution, pH, salt- and protein content. Residual infectivity of the inactivated virus could have disastrous consequences for both laboratory/healthcare personnel and patients. Therefore, the development of inactivation protocols requires careful considerations which we review here.
Highlights
The factors that influence successful inactivation are determined by the inherent characteristics of the virus, the environmental setting and the nature of the intended downstream application
This study demonstrates the complete inactivation of Ebola virus (EBOV) after exposure to a combination of AVL and 0.1% Triton X-100 after incubation for 10 min
As complete inactivation for highly pathogenic viruses is so critical, several studies suggest the combination of inactivating techniques including at least two different modes of action, e.g., one physical and one chemical [8,15,26]
Summary
The factors that influence successful inactivation are determined by the inherent characteristics of the virus, the environmental setting (including the access to equipment and resources) and the nature of the intended downstream application. Serology and vaccine development [25,48,49,50] Effectivity against both RNA and DNA viruses [25]. Acts through an indirect pathway by formation of reactive oxygen species that damage nucleic acids and proteins [1,55,56]. Effect on both viral RNA and DNA [1,27]. Indirect pathway: formation of free radicals from radiolytic cleavage of water (hydroxyl radicals) and oxygen (singlet oxygen) These molecules react with nucleic acids and proteins leaving the virus incapable of replicating [59,60,61,62]. Applications in clinical diagnostics remain to be investigated [67,68]
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