Discovery of DNA viruses in wild-caught mosquitoes using small RNA high throughput sequencing.

<h4>Background</h4>Mosquito-borne infectious diseases pose a severe threat to public health in many areas of the world. Current methods for pathogen detection and surveillance are usually dependent on prior knowledge of the etiologic agents involved. Hence, efficient approaches are requi...

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Autores principales:	Maijuan Ma, Yong Huang, Zhengda Gong, Lu Zhuang, Cun Li, Hong Yang, Yigang Tong, Wei Liu, Wuchun Cao
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3618c7419b4d4cbdbcbfd8b6881cf66b
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spelling	oai:doaj.org-article:3618c7419b4d4cbdbcbfd8b6881cf66b2021-11-04T06:08:13ZDiscovery of DNA viruses in wild-caught mosquitoes using small RNA high throughput sequencing.1932-620310.1371/journal.pone.0024758https://doaj.org/article/3618c7419b4d4cbdbcbfd8b6881cf66b2011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21949749/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Mosquito-borne infectious diseases pose a severe threat to public health in many areas of the world. Current methods for pathogen detection and surveillance are usually dependent on prior knowledge of the etiologic agents involved. Hence, efficient approaches are required for screening wild mosquito populations to detect known and unknown pathogens.<h4>Methodology/principal findings</h4>In this study, we explored the use of Next Generation Sequencing to identify viral agents in wild-caught mosquitoes. We extracted total RNA from different mosquito species from South China. Small 18-30 bp length RNA molecules were purified, reverse-transcribed into cDNA and sequenced using Illumina GAIIx instrumentation. Bioinformatic analyses to identify putative viral agents were conducted and the results confirmed by PCR. We identified a non-enveloped single-stranded DNA densovirus in the wild-caught Culex pipiens molestus mosquitoes. The majority of the viral transcripts (.>80% of the region) were covered by the small viral RNAs, with a few peaks of very high coverage obtained. The +/- strand sequence ratio of the small RNAs was approximately 7∶1, indicating that the molecules were mainly derived from the viral RNA transcripts. The small viral RNAs overlapped, enabling contig assembly of the viral genome sequence. We identified some small RNAs in the reverse repeat regions of the viral 5'- and 3' -untranslated regions where no transcripts were expected.<h4>Conclusions/significance</h4>Our results demonstrate for the first time that high throughput sequencing of small RNA is feasible for identifying viral agents in wild-caught mosquitoes. Our results show that it is possible to detect DNA viruses by sequencing the small RNAs obtained from insects, although the underlying mechanism of small viral RNA biogenesis is unclear. Our data and those of other researchers show that high throughput small RNA sequencing can be used for pathogen surveillance in wild mosquito vectors.Maijuan MaYong HuangZhengda GongLu ZhuangCun LiHong YangYigang TongWei LiuWuchun CaoPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 9, p e24758 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Maijuan Ma Yong Huang Zhengda Gong Lu Zhuang Cun Li Hong Yang Yigang Tong Wei Liu Wuchun Cao Discovery of DNA viruses in wild-caught mosquitoes using small RNA high throughput sequencing.
description	<h4>Background</h4>Mosquito-borne infectious diseases pose a severe threat to public health in many areas of the world. Current methods for pathogen detection and surveillance are usually dependent on prior knowledge of the etiologic agents involved. Hence, efficient approaches are required for screening wild mosquito populations to detect known and unknown pathogens.<h4>Methodology/principal findings</h4>In this study, we explored the use of Next Generation Sequencing to identify viral agents in wild-caught mosquitoes. We extracted total RNA from different mosquito species from South China. Small 18-30 bp length RNA molecules were purified, reverse-transcribed into cDNA and sequenced using Illumina GAIIx instrumentation. Bioinformatic analyses to identify putative viral agents were conducted and the results confirmed by PCR. We identified a non-enveloped single-stranded DNA densovirus in the wild-caught Culex pipiens molestus mosquitoes. The majority of the viral transcripts (.>80% of the region) were covered by the small viral RNAs, with a few peaks of very high coverage obtained. The +/- strand sequence ratio of the small RNAs was approximately 7∶1, indicating that the molecules were mainly derived from the viral RNA transcripts. The small viral RNAs overlapped, enabling contig assembly of the viral genome sequence. We identified some small RNAs in the reverse repeat regions of the viral 5'- and 3' -untranslated regions where no transcripts were expected.<h4>Conclusions/significance</h4>Our results demonstrate for the first time that high throughput sequencing of small RNA is feasible for identifying viral agents in wild-caught mosquitoes. Our results show that it is possible to detect DNA viruses by sequencing the small RNAs obtained from insects, although the underlying mechanism of small viral RNA biogenesis is unclear. Our data and those of other researchers show that high throughput small RNA sequencing can be used for pathogen surveillance in wild mosquito vectors.
format	article
author	Maijuan Ma Yong Huang Zhengda Gong Lu Zhuang Cun Li Hong Yang Yigang Tong Wei Liu Wuchun Cao
author_facet	Maijuan Ma Yong Huang Zhengda Gong Lu Zhuang Cun Li Hong Yang Yigang Tong Wei Liu Wuchun Cao
author_sort	Maijuan Ma
title	Discovery of DNA viruses in wild-caught mosquitoes using small RNA high throughput sequencing.
title_short	Discovery of DNA viruses in wild-caught mosquitoes using small RNA high throughput sequencing.
title_full	Discovery of DNA viruses in wild-caught mosquitoes using small RNA high throughput sequencing.
title_fullStr	Discovery of DNA viruses in wild-caught mosquitoes using small RNA high throughput sequencing.
title_full_unstemmed	Discovery of DNA viruses in wild-caught mosquitoes using small RNA high throughput sequencing.
title_sort	discovery of dna viruses in wild-caught mosquitoes using small rna high throughput sequencing.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/3618c7419b4d4cbdbcbfd8b6881cf66b
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Discovery of DNA viruses in wild-caught mosquitoes using small RNA high throughput sequencing.

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