dsRNA-Seq: Identification of Viral Infection by Purifying and Sequencing dsRNA. Academic Article uri icon

Overview

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 RNA viruses produce double-stranded RNA (dsRNA) while replicating. Purifying and sequencing dsRNA from the total RNA isolated from infected tissue allowed us to recover dsRNA virus sequences and replicated sequences from single-stranded RNA (ssRNA) viruses. We refer to this approach as dsRNA-Seq. By assembling dsRNA sequences into contigs we identified full length or partial RNA viral genomes 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 don't have sequenced genomes and/or commercially available rRNA depletion reagents. In addition, a significant advantage of this method is the ability to identify replicated viral sequences of ssRNA viruses, which is useful for distinguishing infectious viral agents from potential noninfectious viral particles or contaminants.

publication date

  • October 14, 2019

Research

keywords

  • High-Throughput Nucleotide Sequencing
  • RNA Virus Infections
  • RNA Viruses
  • RNA, Double-Stranded

Identity

PubMed Central ID

  • PMC6832592

Scopus Document Identifier

  • 85073440892

Digital Object Identifier (DOI)

  • 10.3390/v11100943

PubMed ID

  • 31615058

Additional Document Info

volume

  • 11

issue

  • 10