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Use of S1 nuclease in deep sequencing for detection of double-stranded RNA viruses

Metagenomic approach using next-generation DNA sequencing has facilitated the detection of many pathogenic viruses from fecal samples. However, in many cases, majority of the detected sequences originate from the host genome and bacterial flora in the gut. Here, to improve efficiency of the detectio...

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Autores principales: SHIMADA, Saya, NAGAI, Makoto, MORIYAMA, Hiromitsu, FUKUHARA, Toshiyuki, KOYAMA, Satoshi, OMATSU, Tsutomu, FURUYA, Tetsuya, SHIRAI, Junsuke, MIZUTANI, Tetsuya
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Japanese Society of Veterinary Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4591160/
https://www.ncbi.nlm.nih.gov/pubmed/25843154
http://dx.doi.org/10.1292/jvms.14-0607
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author SHIMADA, Saya
NAGAI, Makoto
MORIYAMA, Hiromitsu
FUKUHARA, Toshiyuki
KOYAMA, Satoshi
OMATSU, Tsutomu
FURUYA, Tetsuya
SHIRAI, Junsuke
MIZUTANI, Tetsuya
author_facet SHIMADA, Saya
NAGAI, Makoto
MORIYAMA, Hiromitsu
FUKUHARA, Toshiyuki
KOYAMA, Satoshi
OMATSU, Tsutomu
FURUYA, Tetsuya
SHIRAI, Junsuke
MIZUTANI, Tetsuya
author_sort SHIMADA, Saya
collection PubMed
description Metagenomic approach using next-generation DNA sequencing has facilitated the detection of many pathogenic viruses from fecal samples. However, in many cases, majority of the detected sequences originate from the host genome and bacterial flora in the gut. Here, to improve efficiency of the detection of double-stranded (ds) RNA viruses from samples, we evaluated the applicability of S1 nuclease on deep sequencing. Treating total RNA with S1 nuclease resulted in 1.5–28.4- and 10.1–208.9-fold increases in sequence reads of group A rotavirus in fecal and viral culture samples, respectively. Moreover, increasing coverage of mapping to reference sequences allowed for sufficient genotyping using analytical software. These results suggest that library construction using S1 nuclease is useful for deep sequencing in the detection of dsRNA viruses.
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spelling pubmed-45911602015-10-02 Use of S1 nuclease in deep sequencing for detection of double-stranded RNA viruses SHIMADA, Saya NAGAI, Makoto MORIYAMA, Hiromitsu FUKUHARA, Toshiyuki KOYAMA, Satoshi OMATSU, Tsutomu FURUYA, Tetsuya SHIRAI, Junsuke MIZUTANI, Tetsuya J Vet Med Sci Virology Metagenomic approach using next-generation DNA sequencing has facilitated the detection of many pathogenic viruses from fecal samples. However, in many cases, majority of the detected sequences originate from the host genome and bacterial flora in the gut. Here, to improve efficiency of the detection of double-stranded (ds) RNA viruses from samples, we evaluated the applicability of S1 nuclease on deep sequencing. Treating total RNA with S1 nuclease resulted in 1.5–28.4- and 10.1–208.9-fold increases in sequence reads of group A rotavirus in fecal and viral culture samples, respectively. Moreover, increasing coverage of mapping to reference sequences allowed for sufficient genotyping using analytical software. These results suggest that library construction using S1 nuclease is useful for deep sequencing in the detection of dsRNA viruses. The Japanese Society of Veterinary Science 2015-04-06 2015-09 /pmc/articles/PMC4591160/ /pubmed/25843154 http://dx.doi.org/10.1292/jvms.14-0607 Text en ©2015 The Japanese Society of Veterinary Science http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License.
spellingShingle Virology
SHIMADA, Saya
NAGAI, Makoto
MORIYAMA, Hiromitsu
FUKUHARA, Toshiyuki
KOYAMA, Satoshi
OMATSU, Tsutomu
FURUYA, Tetsuya
SHIRAI, Junsuke
MIZUTANI, Tetsuya
Use of S1 nuclease in deep sequencing for detection of double-stranded RNA viruses
title Use of S1 nuclease in deep sequencing for detection of double-stranded RNA viruses
title_full Use of S1 nuclease in deep sequencing for detection of double-stranded RNA viruses
title_fullStr Use of S1 nuclease in deep sequencing for detection of double-stranded RNA viruses
title_full_unstemmed Use of S1 nuclease in deep sequencing for detection of double-stranded RNA viruses
title_short Use of S1 nuclease in deep sequencing for detection of double-stranded RNA viruses
title_sort use of s1 nuclease in deep sequencing for detection of double-stranded rna viruses
topic Virology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4591160/
https://www.ncbi.nlm.nih.gov/pubmed/25843154
http://dx.doi.org/10.1292/jvms.14-0607
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