Methods of Inactivation of Highly Pathogenic Viruses for Molecular, Serology or Vaccine Development Purposes

Abstract

The handling of highly pathogenic viruses, whether for diagnostic or research purposes, often requires an activation 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.

A minimally manipulated preservation and virus inactivation method for amnion/chorion

“…The commonly using virus inactivation technologies include the treatment with gamma irradiation, heat or UV-radiation; the treatment with photoactive compounds such as methylene blue, psoralens, or 1.5-iodonaphtyl azide; the treatment with ionic or non-ionic detergents such as Tween-20/Tween-80, Triton X-100 or sodium dodecyl sulfate and with solvents such as alcohol and acetone ( Elveborg et al, 2022 ). The combination of simple techniques such as heat, UV-radiation, or detergents are often sufficient for virus inactivation but the efficiency depends on the infectious pathogens and the nature of the biomedical products.…”

Contributions of vibrational spectroscopy to virology: A review

“…Viruses and virus containing samples can be inactivated for safe handling under normal laboratory conditions (BSL-1) [48] , by for example chemical [49] , [50] , heat [51] or radiation treatment [52] .…”

Influence of SARS-CoV-2 inactivation by different chemical reagents on the humoral response evaluated in a murine model

“…According to Rodgers et al, 1985 , other inactivation procedures, such as the application of heat, alcohol, or radiation, can impair the structural characteristics of viruses, not being suitable for applications that require such viral preservation. Nevertheless, Gamma irradiation can also be used to inactivate viruses, although multiple influencing factors must be considered, such as solute protein content, virus concentration, temperature and the volume of air, possibly related to the amount of oxygen free radicals produced in the reaction (Elliott et al, 2016; Hume et al, 2016 ; Elveborg et al, 2022 ). Furthermore, considering that SARS-CoV-2 is classified as a Risk Group 3 biological agent, high-titer virus stocks require biosafety level 3 (BSL-3) facilities for its handling ( Yeh et al, 2021 ) but gamma irradiators are not located within these laboratories due to a multitude of practical reasons ( Hume et al, 2016 ).…”

“…Other operational disadvantages of using gamma irradiation include the need to work with radioactive materials and to remain in compliance with the increasing regulatory requirements naturally desirable to minimize health and environmental risks (IAEA, 2016). In this sense, there are still some concerns regarding biosafety, reduced antigenicity and the fact that the equipment is too bulky and expensive for some settings ( Elveborg et al, 2022 ).…”