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  4. Comparison of Three Viral Nucleic Acid Preamplification Pipelines for Sewage Viral Metagenomics
 
research article

Comparison of Three Viral Nucleic Acid Preamplification Pipelines for Sewage Viral Metagenomics

Fernandez Cassi, Xavier  
•
Kohn, Tamar  
2024
Food and Environmental Virology

Viral metagenomics is a useful tool for detecting multiple human viruses in urban sewage. However, more refined protocols are required for its effective use in disease surveillance. In this study, we investigated the performance of three different preamplification pipelines (specific to RNA viruses, DNA viruses or both) for viral genome sequencing using spiked-in Phosphate Buffered Saline and sewage samples containing known concentrations of viruses. We found that compared to the pipeline targeting all genome types, the RNA pipeline performed better in detecting RNA viruses in both spiked and unspiked sewage samples, allowing the detection of various mammalian viruses including members from the Reoviridae, Picornaviridae, Astroviridae and Caliciviridae. However, the DNA-specific pipeline did not improve the detection of mammalian DNA viruses. We also measured viral recovery by quantitative reverse transcription polymerase chain reaction and assessed the impact of genetic background (non-viral genetic material) on viral coverage. Our results indicate that viral recoveries were generally lower in sewage (average of 11.0%) and higher in Phosphate Buffered Saline (average of 23.4%) for most viruses. Additionally, spiked-in viruses showed lower genome coverage in sewage, demonstrating the negative effect of genetic background on sequencing. Finally, correlation analysis revealed a relationship between virus concentration and genome normalized reads per million, indicating that viral metagenomic sequencing can be semiquantitative.

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Type
research article
DOI
10.1007/s12560-024-09594-3
Author(s)
Fernandez Cassi, Xavier  
Kohn, Tamar  
Date Issued

2024

Published in
Food and Environmental Virology
Subjects

Viral metagenomics

•

Sewage monitoring

•

Environmental surveillance

•

Enteric viruses

•

Waterborne pathogens

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
LEV  
FunderGrant Number

H2020

754462

RelationURL/DOI

IsSupplementedBy

https://www.ncbi.nlm.nih.gov/bioproject/948647
Available on Infoscience
April 23, 2024
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/207467
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