Evaluation of metatranscriptomic sequencing protocols to obtain full-length RNA virus genomes from mammalian tissues

Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081766%3A_____%2F25%3A00636787" target="_blank" >RIV/68081766:_____/25:00636787 - isvavai.cz</a>
Result on the web
<a href="https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0324537" target="_blank" >https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0324537</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1371/journal.pone.0324537" target="_blank" >10.1371/journal.pone.0324537</a>

Result language
angličtina
Original language name
Evaluation of metatranscriptomic sequencing protocols to obtain full-length RNA virus genomes from mammalian tissues
Original language description
High-throughput sequencing technologies have advanced RNA virus genomics, but recovering viral genomes from mammalian tissues remains challenging due to the predominance of host RNA. We evaluated two metatranscriptomic workflows to address these challenges. Our results demonstrate that the methods differed significantly in performance, with Method B achieving a 5-fold increase in RNA yield and improved RNA integrity over Method A. These differences resulted in the recovery of 4 complete hepacivirus genomes with Method B compared to fragmented or incomplete genomes with Method A. Additionally, Method B's library preparation workflow, incorporating rRNA depletion, enhanced viral genome recovery by reducing host RNA background. Our novel approach integrates an optimized RNA purification protocol with a customized bioinformatics strategy for improved viral genome recovery. Overall, our findings highlight the critical role of optimized homogenization, RNA purification, and library preparation in metatranscriptomic workflows, facilitating the more effective RNA virus genome recovery from complex mammalian tissues.
Czech name
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Czech description
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Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
—
OECD FORD branch
10613 - Zoology

Project
<a href="/en/project/GA22-32394S" target="_blank" >GA22-32394S: Viromics of the European house mouse hybrid zone: a model to understand how viruses cross host species barriers</a><br>
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year
2025
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

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