Development of Conventional Multiplex PCR Assays for the Identification of 21 West Palaearctic Biting Midge Taxa (Diptera: Ceratopogonidae) Belonging to the Culicoides Subgenus Culicoides, including Recently Discovered Species and Genetic Variants

In 2006 and 2011, two biting-midge-borne arboviruses of high veterinary importance emerged for the first time in Central Europe: bluetongue virus (BTV) Schmallenberg (SBV). Members native Obsoletus Pulicaris Groups were soon identified as potential vectors. However, despite several years extensive taxonomic research on these groups, correct species identification differentiation from closely related are still challenging due to isomorphic features, existence cryptic obsolete PCR assays. At present, 17 valid West Palaearctic biting midge Culicoides subgenus Culicoides, including Group, known, additional genetic variants have been described. For many them, no tests available, their roles disease transmission remained unknown. this study, 465 GenBank DNA sequence entries mitochondrial cytochrome c oxidase subunit I (COI) gene used design primers specific markers 21 taxa Culicoides. During validation with field-collected midges synthetic genotypes not available field, all detected target taxa, while few showed cross-reactions. Our results indicate great new assays clearly demonstrate suitability COI an excellent marker different