Tissue-Based Universal Virus Detection (TUViD-VM) Protocol for Viral Metagenomics.

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.