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
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Date
2014
Authors
Garros, ClaireBalenghien, 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
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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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Bibliographic citation
Parasites & Vectors, 7:223, p. 1-9
AGROVOC subjects
CulicoidesIdentificación
PCR