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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- 作者：Claire Garros (3) (4)
Thomas Balenghien (3) (4)
Simon Carpenter (5)
Jean-Claude Del茅colle (6)
Rudy Meiswinkel (7)
Aur茅lie P茅darrieu (3) (4)
Ignace Rakotoarivony (3) (4)
Laetitia Gard猫s (3) (4)
Nick Golding (5)
James Barber (5)
Miguel Miranda (8)
David Borr脿s Borr脿s (8)
Maria Goffredo (9)
Federica Monaco (9)
Nonito Pag猫s (10)
Soufien Sghaier (11)
Salah Hammami (11)
Jorge H Calvo (12)
Javier Lucientes (13)
Dirk Geysen (14)
Gill De Deken (14)
Victor Sarto i Monteys (15)
Jan Schwenkenbecher (16)
Helge Kampen (17)
Bernd Hoffmann (17)
Kathrin Lehmann (18)
Doreen Werner (19)
Thierry Baldet (3) (4)
Renaud Lancelot (3) (4)
Catherine C锚tre-Sossah (3) (4)
3. Cirad ; UMR15 CMAEE ; 34398 ; Montpellier ; France
4. INRA ; UMR1309 CMAEE ; 34398 ; Montpellier ; France
5. The Pirbright Institute ; Pirbright ; UK
6. Facult茅 de M茅decine de Strasbourg ; IPPTS ; Strasbourg ; France
7. CIDC ; Central Institute for Animal diseases ; Lelystad ; Netherlands
8. Laboratory of Zoology ; UIB ; University of Balearic Islands ; Palma de Mallorca ; Spain
9. IZS ; Istituto Zooprofilatico Sperimentale dell鈥橝bruzzo e del Molise 鈥淕 Caporale鈥? Teramo ; Italy
10. Centre de Recerca en Sanitat Animal (CReSA) ; UAB-IRTA ; Campus de la Universitat Aut貌noma de Barcelona ; Bellaterra (Cerdanyola del Vall猫s) ; Spain
11. IRVT ; Institut de la Recherche V茅t茅rinaire de Tunisie ; Universit茅 Tunis El Manar ; Tunis ; Tunisia
12. CITA-ARAID ; Centro de Investigacion y Tecnologia Agroalimentaria de Aragon ; Zaragoza ; Spain
13. UZ ; Facultad de Veterinaria ; Universidad de Zaragoza ; Zaragoza ; Spain
14. Institute of Tropical Medicine ; Antwerp ; Belgium
15. Institut de Ci猫ncia i Tecnologia Ambientals (ICTA) ; Bellaterra ; Spain
16. IBES ; Institute of Biological and Environmental Sciences ; University of Aberdeen ; Aberdeen ; Scotland
17. FLI ; Friedrich-Loeffler Institut ; Greifswald-Insel Riems ; Germany
18. Humboldt-Universit盲t zu Berlin ; Cytogenetics ; Chausseestr 117 ; Berlin ; Germany
19. Leibniz Centre for Agricultural Landscape Research (ZALF) ; M眉ncheberg ; Germany - 关键词：Culicoides ; Subgenus Avaritia ; Obsoletus group ; Molecular identification
- 刊名：Parasites & Vectors
- 出版年：2014
- 出版时间：December 2014
- 年：2014
- 卷：7
- 期：1
- 全文大小：225 KB
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- 出版者：BioMed Central
- ISSN：1756-3305
文摘
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.