Multi-gene detection and identification of mosquito-borne RNA viruses using an oligonucleotide microarray.

<h4>Background</h4>Arthropod-borne viruses are important emerging pathogens world-wide. Viruses transmitted by mosquitoes, such as dengue, yellow fever, and Japanese encephalitis viruses, infect hundreds of millions of people and animals each year. Global surveillance of these viruses in...

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Autores principales:	Nathan D Grubaugh, Scott S McMenamy, Michael J Turell, John S Lee
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270
Acceso en línea:	https://doaj.org/article/0dd3d1e694cd4647ad3594b5a8dfbb59
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spelling	oai:doaj.org-article:0dd3d1e694cd4647ad3594b5a8dfbb592021-11-18T09:17:01ZMulti-gene detection and identification of mosquito-borne RNA viruses using an oligonucleotide microarray.1935-27271935-273510.1371/journal.pntd.0002349https://doaj.org/article/0dd3d1e694cd4647ad3594b5a8dfbb592013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23967358/?tool=EBIhttps://doaj.org/toc/1935-2727https://doaj.org/toc/1935-2735<h4>Background</h4>Arthropod-borne viruses are important emerging pathogens world-wide. Viruses transmitted by mosquitoes, such as dengue, yellow fever, and Japanese encephalitis viruses, infect hundreds of millions of people and animals each year. Global surveillance of these viruses in mosquito vectors using molecular based assays is critical for prevention and control of the associated diseases. Here, we report an oligonucleotide DNA microarray design, termed ArboChip5.1, for multi-gene detection and identification of mosquito-borne RNA viruses from the genera Flavivirus (family Flaviviridae), Alphavirus (Togaviridae), Orthobunyavirus (Bunyaviridae), and Phlebovirus (Bunyaviridae).<h4>Methodology/principal findings</h4>The assay utilizes targeted PCR amplification of three genes from each virus genus for electrochemical detection on a portable, field-tested microarray platform. Fifty-two viruses propagated in cell-culture were used to evaluate the specificity of the PCR primer sets and the ArboChip5.1 microarray capture probes. The microarray detected all of the tested viruses and differentiated between many closely related viruses such as members of the dengue, Japanese encephalitis, and Semliki Forest virus clades. Laboratory infected mosquitoes were used to simulate field samples and to determine the limits of detection. Additionally, we identified dengue virus type 3, Japanese encephalitis virus, Tembusu virus, Culex flavivirus, and a Quang Binh-like virus from mosquitoes collected in Thailand in 2011 and 2012.<h4>Conclusions/significance</h4>We demonstrated that the described assay can be utilized in a comprehensive field surveillance program by the broad-range amplification and specific identification of arboviruses from infected mosquitoes. Furthermore, the microarray platform can be deployed in the field and viral RNA extraction to data analysis can occur in as little as 12 h. The information derived from the ArboChip5.1 microarray can help to establish public health priorities, detect disease outbreaks, and evaluate control programs.Nathan D GrubaughScott S McMenamyMichael J TurellJohn S LeePublic Library of Science (PLoS)articleArctic medicine. Tropical medicineRC955-962Public aspects of medicineRA1-1270ENPLoS Neglected Tropical Diseases, Vol 7, Iss 8, p e2349 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270
spellingShingle	Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Nathan D Grubaugh Scott S McMenamy Michael J Turell John S Lee Multi-gene detection and identification of mosquito-borne RNA viruses using an oligonucleotide microarray.
description	<h4>Background</h4>Arthropod-borne viruses are important emerging pathogens world-wide. Viruses transmitted by mosquitoes, such as dengue, yellow fever, and Japanese encephalitis viruses, infect hundreds of millions of people and animals each year. Global surveillance of these viruses in mosquito vectors using molecular based assays is critical for prevention and control of the associated diseases. Here, we report an oligonucleotide DNA microarray design, termed ArboChip5.1, for multi-gene detection and identification of mosquito-borne RNA viruses from the genera Flavivirus (family Flaviviridae), Alphavirus (Togaviridae), Orthobunyavirus (Bunyaviridae), and Phlebovirus (Bunyaviridae).<h4>Methodology/principal findings</h4>The assay utilizes targeted PCR amplification of three genes from each virus genus for electrochemical detection on a portable, field-tested microarray platform. Fifty-two viruses propagated in cell-culture were used to evaluate the specificity of the PCR primer sets and the ArboChip5.1 microarray capture probes. The microarray detected all of the tested viruses and differentiated between many closely related viruses such as members of the dengue, Japanese encephalitis, and Semliki Forest virus clades. Laboratory infected mosquitoes were used to simulate field samples and to determine the limits of detection. Additionally, we identified dengue virus type 3, Japanese encephalitis virus, Tembusu virus, Culex flavivirus, and a Quang Binh-like virus from mosquitoes collected in Thailand in 2011 and 2012.<h4>Conclusions/significance</h4>We demonstrated that the described assay can be utilized in a comprehensive field surveillance program by the broad-range amplification and specific identification of arboviruses from infected mosquitoes. Furthermore, the microarray platform can be deployed in the field and viral RNA extraction to data analysis can occur in as little as 12 h. The information derived from the ArboChip5.1 microarray can help to establish public health priorities, detect disease outbreaks, and evaluate control programs.
format	article
author	Nathan D Grubaugh Scott S McMenamy Michael J Turell John S Lee
author_facet	Nathan D Grubaugh Scott S McMenamy Michael J Turell John S Lee
author_sort	Nathan D Grubaugh
title	Multi-gene detection and identification of mosquito-borne RNA viruses using an oligonucleotide microarray.
title_short	Multi-gene detection and identification of mosquito-borne RNA viruses using an oligonucleotide microarray.
title_full	Multi-gene detection and identification of mosquito-borne RNA viruses using an oligonucleotide microarray.
title_fullStr	Multi-gene detection and identification of mosquito-borne RNA viruses using an oligonucleotide microarray.
title_full_unstemmed	Multi-gene detection and identification of mosquito-borne RNA viruses using an oligonucleotide microarray.
title_sort	multi-gene detection and identification of mosquito-borne rna viruses using an oligonucleotide microarray.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/0dd3d1e694cd4647ad3594b5a8dfbb59
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Multi-gene detection and identification of mosquito-borne RNA viruses using an oligonucleotide microarray.

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