Abstract

RNA viruses are a major source of emerging and re-emerging infectious diseases around the world. We developed a method to identify RNA viruses that is based on the fact that all RNA viruses produce dsRNA while replicating. Purifying and sequencing dsRNA from total RNA isolated from infected tissue allowed us to recover replicating viral sequences. We refer to this approach as dsRNA-Seq. By assembling dsRNA sequences into contigs we identified full length RNA viruses of varying genome types infecting mammalian culture samples, identified a known viral disease agent in laboratory infected mice, and successfully detected naturally occurring RNA viral infections in reptiles. Here we show that dsRNA-Seq is a preferable method for identifying viruses in organisms that do not have sequenced genomes and/or commercially available rRNA depletion reagents. Similar to other metagenomic strategies, dsRNA-Seq has the potential to identify unknown viral disease agents that share little to no similarity to known viruses. However, the significant advantage of this method is the ability to identify replicated viral sequences, which is useful for distinguishing infectious viral agents from potential noninfectious viral particles or contaminants.

Details

Title
dsRNA-Seq: Identification of viral infection by purifying and sequencing dsRNA
Author
Decker, Carolyn J; Steiner, Halley R; Hoon-Hanks, Laura L; Morrison, James H; Haist, Kelsey C; Stabell, Alex C; Poeschla, Eric M; Morrison, Thomas E; Stenglein, Mark D; Sawyer, Sara L; Parker, Roy
University/institution
Cold Spring Harbor Laboratory Press
Section
New Results
Publication year
2019
Publication date
Aug 19, 2019
Publisher
Cold Spring Harbor Laboratory Press
ISSN
2692-8205
Source type
Working Paper
Language of publication
English
ProQuest document ID
2275694928
Copyright
© 2019. This article is published under http://creativecommons.org/licenses/by/4.0/ (“the License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.