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

Claire Garros, Thomas Balenghien, Simon Carpenter, Jean Claude Delécolle, Rudy Meiswinkel, Aurélie Pédarrieu, Ignace Rakotoarivony, Laetitia Gardès, Nick Golding, James Barber, Miguel Miranda, David Borràs Borràs, Maria Goffredo, Federica Monaco, Nonito Pagès, Soufien Sghaier, Salah Hammami, Jorge H. Calvo, Javier Lucientes, Dirk GeysenGill De Deken, Victor Sarto I Monteys, Jan Schwenkenbecher, Helge Kampen, Bernd Hoffmann, Kathrin Lehmann, Doreen Werner, Thierry Baldet, Renaud Lancelot, Catherine Cêtre-Sossah

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    13 Citations (Scopus)

    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. © 2014 Garros et al.; licensee BioMed Central Ltd.
    Original languageEnglish
    Article number223
    JournalParasites and Vectors
    Volume7
    Issue number1
    DOIs
    Publication statusPublished - 14 May 2014

    Keywords

    • Culicoides
    • Molecular identification
    • Obsoletus group
    • Subgenus Avaritia

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