Abstract
The protocol for tissue-based universal virus detection (TUViD-VM) was developed to allow for the detection of yet unknown viruses from infected tissues. Tissues are a challenging source when it comes to virus detection. The very rapid degradation rate of tissue and the high level of host nucleic acids are the biggest challenges when examining tissues. However, to overcome these challenges, we had to decrease the amount of host nucleic acids rigorously in order to increase the amount of viral nucleic acids. We compared modern and common virological approaches to find the optimal conditions. The final TUViD-VM protocol was extensively validated by using real-time PCR and next-generation sequencing. We could increase the amount of detectable virus nucleic acids and improved the detection of viruses <75,000-fold compared with other approaches tested. The TUViD-VM protocol can be used in metagenomic and virome studies to increase the likelihood of detecting viruses from any biological source. The protocol has been widely used to detect viruses in different tissues with great success. Here we provide the bench-top protocol version of the TUViD-VM protocol.
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References
Kohl C, Brinkmann A, Dabrowski PW et al (2015) Protocol for metagenomic virus detection in clinical specimens. Emerg Infect Dis 21(1):48–57
Stang A, Korn K, Wildner O et al (2005) Characterization of virus isolates by particle-associated nucleic acid PCR. J Clin Microbiol 43(2):716–720
Kohl C, Wegener M, Nitsche A et al (2017) Use of RNALater® preservation for Virome sequencing in outbreak settings. Front Microbiol 8:1888
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The authors are grateful to Ursula Erikli for copy-editing.
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Kohl, C., Kurth, A. (2018). Tissue-Based Universal Virus Detection (TUViD-VM) Protocol for Viral Metagenomics. In: Moya, A., Pérez Brocal, V. (eds) The Human Virome. Methods in Molecular Biology, vol 1838. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8682-8_2
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DOI: https://doi.org/10.1007/978-1-4939-8682-8_2
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8681-1
Online ISBN: 978-1-4939-8682-8
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