Implication of cell culture methods and biases on UV inactivation of viruses

ElsevierVolume 309, November 2022, 114610Journal of Virological MethodsHighlights•

Potential artifacts may be caused by interactions between UV-C and culture media.

Reactive oxygen and nitrogen species form in virus culture media exposed to 265 nm.

Inadequate virus media may cause a bias for the UV-C required for virus inactivation.

Abstract

Inactivation of human respiratory viruses in air and on surfaces is important to control their spread. Exposure to germicidal ultraviolet (UV-C) light damages viral nucleic acid rendering them non-infectious. Most of the recent viral inactivation studies have not considered potential artifacts caused by interactions between UV-C light and culture media used to suspend and deposit virus on surfaces. We show that the reactive oxygen and nitrogen species (ROS and RNS) form when commonly used virus culture media is exposed to 265 nm irradiation from light emitting diodes (LEDs) at UV-C doses (4 or 40 mJ/cm2) commonly considered to achieve multiple log-inactivation of virus. Surface viral inactivation values were enhanced from 0.49 to 2.92 log10 of viruses in DMEM, EMEM or EMEM-F as compared to absence of culture media (only suspended in Tris-buffer). The mechanisms responsible for the enhanced surface inactivate is hypothesized to involve photo-activation of vitamins and dyes present in the culture media, deposited with the virus on surfaces to be disinfected, which produce ROS and RNS. Given the rapidly growing research and commercial markets for UV-C disinfecting devices, there is a need to establish surface disinfecting protocols that avoid viral inactivation enhancement artifacts associated with selection and use of common cell culture media in the presence of UV-C light. This study addresses this weak link in the literature and highlights that inadequate selection of virus suspension media may cause a bias (i.e., over-estimation) for the UV-C dosages required for virus inactivation on surfaces.

Keywords

Ultraviolet light (UV)

Light emitting diode (LED)

Virus inactivation

Reactive oxygen species

Reactive nitrogen species

UV disinfection

Human coronavirus (HuCoV)

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© 2022 Published by Elsevier B.V.

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