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

Kód výsledku v 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>

Výsledek na webu

<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>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10613 - Zoology

Projekt

<a href="/cs/project/GA22-32394S" target="_blank" >GA22-32394S: Viromika myši domácí na hybridní zóně v Evropě: model pro studium šíření virů a překonávání druhových bariér</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2025

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

PLoS ONE

ISSN

1932-6203

e-ISSN

1932-6203

Svazek periodika

20

Číslo periodika v rámci svazku

5

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

15

Strana od-do

e0324537

Kód UT WoS článku

001499528700004

EID výsledku v databázi Scopus

2-s2.0-105007049984