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The causative agent of DF is dengue virus (genus <jats:italic>Flavivirus<\/jats:italic>) that comprises four distinct serotypes (DENV-1 to DENV-4). Fluorescence <jats:italic>in situ<\/jats:italic> hybridization (FISH) has been used successfully to detect pathogenic agents, but has not been implemented in detecting DENV. To improve our understanding of DENV infection and dissemination in host tissues, we designed specific probes to detect DENV in FISH assays.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Methods<\/jats:title>\n            <jats:p>Oligonucleotide probes were designed to hybridize with RNA from the broadest range of DENV isolates belonging to the four serotypes, but not to the closest <jats:italic>Flavivirus<\/jats:italic> genomes. Three probes that fit the criteria defined for FISH experiments were selected, targeting both coding and non-coding regions of the DENV genome. These probes were tested in FISH assays against the dengue vector <jats:italic>Aedes albopictus<\/jats:italic> (<jats:italic>Diptera<\/jats:italic>: <jats:italic>Culicidae<\/jats:italic>). The FISH experiments were led <jats:italic>in vitro<\/jats:italic> using the C6\/36 cell line, and <jats:italic>in vivo<\/jats:italic> against dissected salivary glands, with epifluorescence and confocal microscopy.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>The three 60-nt oligonucleotides probes DENV-Probe A, B and C cover a broad range of DENV isolates from the four serotypes. When the three probes were used together, specific fluorescent signals were observed in C6\/36 infected with each DENV serotypes. No signal was detected in either cells infected with close <jats:italic>Flavivirus<\/jats:italic> members West Nile virus or yellow fever virus. The same protocol was used on salivary glands of <jats:italic>Ae. albopictus<\/jats:italic> fed with a DENV-2 infectious blood-meal which showed positive signals in the lateral lobes of infected samples, with no significant signal in uninfected mosquitoes.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusion<\/jats:title>\n            <jats:p>Based on the FISH technique, we propose a way to design and use oligonucleotide probes to detect arboviruses. Results showed that this method was successfully implemented to specifically detect DENV in a mosquito cell line, as well as in mosquito salivary glands for the DENV-2 serotype. In addition, we emphasize that FISH could be an alternative method to detect arboviruses in host tissues, also offering to circumvent the discontinuity of antibodies used in immunofluorescent assays.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/1756-3305-5-243","type":"journal-article","created":{"date-parts":[[2012,10,31]],"date-time":"2012-10-31T17:17:27Z","timestamp":1351703847000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Detection of dengue group viruses by fluorescence in situ hybridization"],"prefix":"10.1186","volume":"5","author":[{"given":"Vincent","family":"Raquin","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Wannagat","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karima","family":"Zouache","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Catherine","family":"Legras-Lachuer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claire Valiente","family":"Moro","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick","family":"Mavingui","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2012,10,30]]},"reference":[{"key":"733_CR1","doi-asserted-by":"publisher","first-page":"S7","DOI":"10.1038\/nrmicro2460","volume":"8","author":"MG Guzman","year":"2010","unstructured":"Guzman MG, Halstead SB, Artsob H, Buchy P, Farrar J, Gubler DJ, Hunsperger E, Kroeger A, Margolis HS, Mart\u00ednez E, Nathan MB, Pelegrino JL, Simmons C, Yoksan S, Peeling RW: Dengue: a continuing global threat. 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