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.
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.
authors.affiliation: Cirad, UMR15 CMAEE, 34398 Montpellier, France. garros@cirad.fr.
authors.forename: Claire
authors.initials: C
authors.lastname: Garros
authors.forename: Thomas
authors.initials: T
authors.lastname: Balenghien
authors.forename: Simon
authors.initials: S
authors.lastname: Carpenter
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Delécolle
authors.forename: Rudy
authors.initials: R
authors.lastname: Meiswinkel
authors.forename: Aurélie
authors.initials: A
authors.lastname: Pédarrieu
authors.forename: Ignace
authors.initials: I
authors.lastname: Rakotoarivony
authors.forename: Laetitia
authors.initials: L
authors.lastname: Gardès
authors.forename: Nick
authors.initials: N
authors.lastname: Golding
authors.forename: James
authors.initials: J
authors.lastname: Barber
authors.forename: Miguel
authors.initials: M
authors.lastname: Miranda
authors.forename: David Borràs
authors.initials: DB
authors.lastname: Borràs
authors.forename: Maria
authors.initials: M
authors.lastname: Goffredo
authors.forename: Federica
authors.initials: F
authors.lastname: Monaco
authors.forename: Nonito
authors.initials: N
authors.lastname: Pagès
authors.forename: Soufien
authors.initials: S
authors.lastname: Sghaier
authors.forename: Salah
authors.initials: S
authors.lastname: Hammami
authors.forename: Jorge H
authors.initials: JH
authors.lastname: Calvo
authors.forename: Javier
authors.initials: J
authors.lastname: Lucientes
authors.forename: Dirk
authors.initials: D
authors.lastname: Geysen
authors.forename: Gill
authors.initials: G
authors.lastname: De Deken
authors.forename: Victor
authors.initials: V
authors.lastname: Sarto I Monteys
authors.forename: Jan
authors.initials: J
authors.lastname: Schwenkenbecher
authors.forename: Helge
authors.initials: H
authors.lastname: Kampen
authors.forename: Bernd
authors.initials: B
authors.lastname: Hoffmann
authors.forename: Kathrin
authors.initials: K
authors.lastname: Lehmann
authors.forename: Doreen
authors.initials: D
authors.lastname: Werner
authors.forename: Thierry
authors.initials: T
authors.lastname: Baldet
authors.forename: Renaud
authors.initials: R
authors.lastname: Lancelot
authors.forename: Catherine
authors.initials: C
authors.lastname: Cêtre-Sossah
mesh_terms: D000818:Animals; D002519:Ceratopogonidae / Q000145:classification / Q000235:genetics* / Q000502:physiology; D004247:DNA / Q000235:genetics*; D005260:Female; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D004247:DNA