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  4. Differences in viral disinfection mechanisms as revealed by quantitative transfection of Echovirus 11 genomes
 
research article

Differences in viral disinfection mechanisms as revealed by quantitative transfection of Echovirus 11 genomes

Torrey, Jason Robert  
•
von Gunten, Urs  
•
Kohn, Tamar  
May 10, 2019
Applied and Environmental Microbiology

Virus inactivation mechanisms can be elucidated by methods which measure the loss of specific virus functionality (e.g., host attachment, genome internalization, genome replication). Genome functionality is frequently assessed by PCR-based methods, which are indirect and potentially inaccurate; genome damage that affects detection by high-fidelity PCR enzymes may not adversely affect the ability of actual cellular enzymes to produce functional virus. Therefore, we here develop a transfection-based assay to quantitatively determine viral genome functionality by inserting viral RNA into host cells directly to measure their ability to produce new functional viruses from damaged viral genomes. Echovirus 11 was treated with ozone, free chlorine (FC), UV254 or heat and reductions in genome functionality and infectivity were compared. Ozone reduced genome functionality proportionally to infectivity, indicating that genome damage is the main mechanism of virus inactivation. In contrast, FC caused little or no loss of genome functionality compared to infectivity, indicating a larger role for protein damage. For UV254, genome functionality loss accounted for approximately 60% of virus inactivation, with the remainder presumably due to protein damage. Heat treatment resulted in no reduction in genome functionality, in agreement with the understanding that heat inactivation results from capsid damage. Our results indicate that there is a fundamental difference between genome integrity reductions measured by PCR enzymes in previous studies and actual genome functionality (whether the genome can produce virus) post disinfection. Compared to PCR, quantitative transfection assays provide a more realistic picture of actual viral genome functionality and overall inactivation mechanisms during disinfection.

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Type
research article
DOI
10.1128/AEM.00961-19
Author(s)
Torrey, Jason Robert  
von Gunten, Urs  
Kohn, Tamar  
Date Issued

2019-05-10

Published in
Applied and Environmental Microbiology
Volume

85

Issue

14

Start page

e00961

End page

19

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
LEV  
LTQE  
Available on Infoscience
May 11, 2019
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/156389
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