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    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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    Author
    Garros, C; Balenghien, T; Carpenter, S; Delecolle, J-C; Meiswinkel, R; Pedarrieu, A; Rakotoarivony, I; Gardes, L; Golding, N; Barber, J; ...
    Date
    2014-05-14
    Source Title
    Parasites and Vectors
    Publisher
    BMC
    University of Melbourne Author/s
    Golding, Nicholas
    Affiliation
    School of BioSciences
    Metadata
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    Document Type
    Journal Article
    Citations
    Garros, C., Balenghien, T., Carpenter, S., Delecolle, J. -C., Meiswinkel, R., Pedarrieu, A., Rakotoarivony, I., Gardes, L., Golding, N., Barber, J., Miranda, M., Borras Borras, D., Goffredo, M., Monaco, F., Pages, N., Sghaier, S., Hammami, S., Calvo, J. H., Lucientes, J. ,... Cetre-Sossah, C. (2014). 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. PARASITES & VECTORS, 7 (1), https://doi.org/10.1186/1756-3305-7-223.
    Access Status
    Open Access
    URI
    http://hdl.handle.net/11343/258261
    DOI
    10.1186/1756-3305-7-223
    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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