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dsRNA-Seq: Identification of Viral Infection by Purifying and Sequencing dsRNA

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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6832592/
https://www.ncbi.nlm.nih.gov/pubmed/31615058
http://dx.doi.org/10.3390/v11100943
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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
author_facet 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
author_sort Decker, Carolyn J.
collection PubMed
description 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.
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spelling pubmed-68325922019-11-25 dsRNA-Seq: Identification of Viral Infection by Purifying and Sequencing dsRNA 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 Viruses Article 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. MDPI 2019-10-14 /pmc/articles/PMC6832592/ /pubmed/31615058 http://dx.doi.org/10.3390/v11100943 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
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
dsRNA-Seq: Identification of Viral Infection by Purifying and Sequencing dsRNA
title dsRNA-Seq: Identification of Viral Infection by Purifying and Sequencing dsRNA
title_full dsRNA-Seq: Identification of Viral Infection by Purifying and Sequencing dsRNA
title_fullStr dsRNA-Seq: Identification of Viral Infection by Purifying and Sequencing dsRNA
title_full_unstemmed dsRNA-Seq: Identification of Viral Infection by Purifying and Sequencing dsRNA
title_short dsRNA-Seq: Identification of Viral Infection by Purifying and Sequencing dsRNA
title_sort dsrna-seq: identification of viral infection by purifying and sequencing dsrna
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6832592/
https://www.ncbi.nlm.nih.gov/pubmed/31615058
http://dx.doi.org/10.3390/v11100943
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