title: Evaluation of a field-portable DNA microarray platform and nucleic acid amplification strategies for the detection of arboviruses, arthropods, and bloodmeals.
abstract: Highly multiplexed assays, such as microarrays, can benefit arbovirus surveillance by allowing researchers to screen for hundreds of targets at once. We evaluated amplification strategies and the practicality of a portable DNA microarray platform to analyze virus-infected mosquitoes. The prototype microarray design used here targeted the non-structural protein 5, ribosomal RNA, and cytochrome b genes for the detection of flaviviruses, mosquitoes, and bloodmeals, respectively. We identified 13 of 14 flaviviruses from virus inoculated mosquitoes and cultured cells. Additionally, we differentiated between four mosquito genera and eight whole blood samples. The microarray platform was field evaluated in Thailand and successfully identified flaviviruses (Culex flavivirus, dengue-3, and Japanese encephalitis viruses), differentiated between mosquito genera (Aedes, Armigeres, Culex, and Mansonia), and detected mammalian bloodmeals (human and dog). We showed that the microarray platform and amplification strategies described here can be used to discern specific information on a wide variety of viruses and their vectors.
authors.affiliation: Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD 21702, USA. nathan.grubaugh@colostate.edu
authors.forename: Nathan D
authors.initials: ND
authors.lastname: Grubaugh
authors.forename: Lawrence N
authors.initials: LN
authors.lastname: Petz
authors.forename: Vanessa R
authors.initials: VR
authors.lastname: Melanson
authors.forename: Scott S
authors.initials: SS
authors.lastname: McMenamy
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Turell
authors.forename: Lewis S
authors.initials: LS
authors.lastname: Long
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Pisarcik
authors.forename: Ampornpan
authors.initials: A
authors.lastname: Kengluecha
authors.forename: Boonsong
authors.initials: B
authors.lastname: Jaichapor
authors.forename: Monica L
authors.initials: ML
authors.lastname: O'Guinn
authors.forename: John S
authors.initials: JS
authors.lastname: Lee
mesh_terms: D000818:Animals; D001103:Arboviruses / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D001181:Arthropods / Q000821:virology*; D001769:Blood / Q000821:virology*; D019295:Computational Biology; D009033:Culicidae / Q000821:virology; D045303:Cytochromes b / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004285:Dogs; D018397:Equidae; D005416:Flavivirus / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity; D005814:Genes, Viral; D006736:Horses; D006801:Humans; D007303:Insect Vectors / Q000821:virology; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D013785:Thailand; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D004279:DNA, Viral; C096041:NS5 protein, flavivirus; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; D045303:Cytochromes b