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Title: Towards the PCR-based identification of Palaearctic Culicoides biting midges (Diptera: Ceratopogonidae): results from an international ring trial targeting four species of the subgenus Avaritia
Authors: Garros, Claire
Balenghien, Thomas
Carpenter, Simon
Delecolle, Jean Claude
Meiswinkel, Rudy
Pedarrieu, Aurelie
Rakotoarivony, Ignace
Gardes, Laetitia
Colding, Nick
Barber, James
Miranda Chueca, Miguel A.
Borrás Borrás, David
Goffredo, Maria
Monaco, Federica
Pages, Nonito
Sghaier, Soufien
Hammami, Salah
Calvo Lacosta, Jorge Hugo
Lucientes Curdi, Javier
Geysen, Dirk
Deken, Gill de
Sarto i Monteys, Victor
Schwenkenbecher, Jan
Kampen, Helge
Hoffmann, Bernd
Lehmann, Kathrin
Werner, Doreen
Baldet, Thierry
Lancelot, Renaud
Cêtre Sossah, Catherine
Issue Date: 2014
Citation: Parasites & Vectors, 7:223, p. 1-9
Abstract: Background: Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) are biological vectors of internationally important arboviruses. To understand the role of Culicoides in the transmission of these viruses, it is essential to correctly identify the species involved. Within the western Palaearctic region, the main suspected vector species, C. obsoletus, C. scoticus, C. dewulfi and C. chiopterus, have similar wing patterns, which makes it difficult to separate and identify them correctly. Methods: In this study, designed as an inter-laboratory ring trial with twelve partners from Europe and North Africa, we assess four PCR-based assays which are used routinely to differentiate the four species of Culicoides listed above. The assays based on mitochondrial or ribosomal DNA or microarray hybridisation were tested using aliquots of Culicoides DNA (extracted using commercial kits), crude lysates of ground specimens and whole Culicoides (265 individuals), and non-Culicoides Ceratopogonidae (13 individuals) collected from across Europe. Results: A total of 800 molecular assays were implemented. The in-house assays functioned effectively, although specificity and sensitivity varied according to the molecular marker and DNA extraction method used. The Obsoletus group specificity was overall high (95-99%) while the sensitivity varied greatly (59.6-100%). DNA extraction methods impacted the sensitivity of the assays as well as the type of sample used as template for the DNA extraction. Conclusions: The results are discussed in terms of current use of species diagnostic assays and the future development of molecular tools for the rapid differentiation of cryptic Culicoides species.
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ISSN: 1756-3305
Appears in Collections:[DOCIART] Artículos científicos, técnicos y divulgativos

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