TY - JOUR
T1 - Implication of cell culture methods and biases on UV inactivation of viruses
AU - Alum, Absar
AU - Zhao, Zhe
AU - Ersan, Mahmut S.
AU - Mewes, Thorsten
AU - Barnes, Michael
AU - Westerhoff, Paul
AU - Abbaszadegan, Morteza
N1 - Publisher Copyright: © 2022
PY - 2022/11
Y1 - 2022/11
N2 - 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.
AB - 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.
KW - Human coronavirus (HuCoV)
KW - Light emitting diode (LED)
KW - Reactive nitrogen species
KW - Reactive oxygen species
KW - UV disinfection
KW - Ultraviolet light (UV)
KW - Virus inactivation
UR - http://www.scopus.com/inward/record.url?scp=85137305089&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85137305089&partnerID=8YFLogxK
U2 - 10.1016/j.jviromet.2022.114610
DO - 10.1016/j.jviromet.2022.114610
M3 - Article
C2 - 36064127
SN - 0166-0934
VL - 309
JO - Journal of Virological Methods
JF - Journal of Virological Methods
M1 - 114610
ER -