Development of Conventional Multiplex PCR Assays for the Identification of 21 West Palaearctic Biting Midge Taxa (Diptera: Ceratopogonidae) Belonging to the <i>Culicoides</i> Subgenus <i>Culicoides</i>, 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) and Schmallenberg virus (SBV). Members of the native Obsoletus and Pulicaris Groups were soon identified as the potential vectors. However, despite...
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MDPI AG
2023-05-01
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| Online Access: | https://www.mdpi.com/1424-2818/15/6/699 |
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| author | Oliver Dähn Doreen Werner Bruno Mathieu Helge Kampen |
| author_facet | Oliver Dähn Doreen Werner Bruno Mathieu Helge Kampen |
| author_sort | Oliver Dähn |
| collection | DOAJ |
| description | In 2006 and 2011, two biting-midge-borne arboviruses of high veterinary importance emerged for the first time in Central Europe: bluetongue virus (BTV) and Schmallenberg virus (SBV). Members of the native Obsoletus and Pulicaris Groups were soon identified as the potential vectors. However, despite several years of extensive taxonomic research on these groups, correct species identification and differentiation from closely related species are still challenging due to isomorphic features, the existence of cryptic species and obsolete PCR identification assays. At present, 17 valid West Palaearctic biting midge species of the <i>Culicoides</i> subgenus <i>Culicoides</i>, including the Pulicaris Group, are known, and additional genetic variants have been described. For many of them, no identification tests are available, and their roles in disease transmission have remained unknown. In this study, 465 GenBank DNA sequence entries of the mitochondrial cytochrome c oxidase subunit I (COI) gene were used to design PCR primers as specific genetic markers for 21 West Palaearctic biting midge taxa of the <i>Culicoides</i> subgenus <i>Culicoides</i>. During their validation with DNA from field-collected biting midges and synthetic DNA from biting midge genotypes not available from the field, all primers detected their target taxa, while few showed cross-reactions. Our results indicate the great potential of the new primers in PCR assays and clearly demonstrate the suitability of the COI gene as an excellent marker for the identification of different biting midge species and genetic variants of the <i>Culicoides</i> subgenus <i>Culicoides</i>. |
| first_indexed | 2024-03-11T02:35:24Z |
| format | Article |
| id | doaj.art-fbb74fda667c4da08ff962d9a6573053 |
| institution | Directory Open Access Journal |
| issn | 1424-2818 |
| language | English |
| last_indexed | 2024-03-11T02:35:24Z |
| publishDate | 2023-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Diversity |
| spelling | doaj.art-fbb74fda667c4da08ff962d9a65730532023-11-18T10:02:17ZengMDPI AGDiversity1424-28182023-05-0115669910.3390/d15060699Development of Conventional Multiplex PCR Assays for the Identification of 21 West Palaearctic Biting Midge Taxa (Diptera: Ceratopogonidae) Belonging to the <i>Culicoides</i> Subgenus <i>Culicoides</i>, including Recently Discovered Species and Genetic VariantsOliver Dähn0Doreen Werner1Bruno Mathieu2Helge Kampen3Friedrich-Loeffler-Institut (FLI), Federal Research Institute for Animal Health, Südufer 10, 17493 Greifswald, GermanyLeibniz Centre for Agricultural Landscape Research (ZALF), Eberswalder Str. 84, 15374 Müncheberg, GermanyInstitute of Parasitology and Tropical Pathology, Université de Strasbourg, DIHP UR 7292, 67000 Strasbourg, FranceFriedrich-Loeffler-Institut (FLI), Federal Research Institute for Animal Health, Südufer 10, 17493 Greifswald, GermanyIn 2006 and 2011, two biting-midge-borne arboviruses of high veterinary importance emerged for the first time in Central Europe: bluetongue virus (BTV) and Schmallenberg virus (SBV). Members of the native Obsoletus and Pulicaris Groups were soon identified as the potential vectors. However, despite several years of extensive taxonomic research on these groups, correct species identification and differentiation from closely related species are still challenging due to isomorphic features, the existence of cryptic species and obsolete PCR identification assays. At present, 17 valid West Palaearctic biting midge species of the <i>Culicoides</i> subgenus <i>Culicoides</i>, including the Pulicaris Group, are known, and additional genetic variants have been described. For many of them, no identification tests are available, and their roles in disease transmission have remained unknown. In this study, 465 GenBank DNA sequence entries of the mitochondrial cytochrome c oxidase subunit I (COI) gene were used to design PCR primers as specific genetic markers for 21 West Palaearctic biting midge taxa of the <i>Culicoides</i> subgenus <i>Culicoides</i>. During their validation with DNA from field-collected biting midges and synthetic DNA from biting midge genotypes not available from the field, all primers detected their target taxa, while few showed cross-reactions. Our results indicate the great potential of the new primers in PCR assays and clearly demonstrate the suitability of the COI gene as an excellent marker for the identification of different biting midge species and genetic variants of the <i>Culicoides</i> subgenus <i>Culicoides</i>.https://www.mdpi.com/1424-2818/15/6/699<i>Culicoides</i>Pulicaris GroupWest Palaearcticvectorspolymerase chain reaction (PCR)mitochondrial cytochrome c oxidase subunit I (COI) |
| spellingShingle | Oliver Dähn Doreen Werner Bruno Mathieu Helge Kampen Development of Conventional Multiplex PCR Assays for the Identification of 21 West Palaearctic Biting Midge Taxa (Diptera: Ceratopogonidae) Belonging to the <i>Culicoides</i> Subgenus <i>Culicoides</i>, including Recently Discovered Species and Genetic Variants Diversity <i>Culicoides</i> Pulicaris Group West Palaearctic vectors polymerase chain reaction (PCR) mitochondrial cytochrome c oxidase subunit I (COI) |
| title | Development of Conventional Multiplex PCR Assays for the Identification of 21 West Palaearctic Biting Midge Taxa (Diptera: Ceratopogonidae) Belonging to the <i>Culicoides</i> Subgenus <i>Culicoides</i>, including Recently Discovered Species and Genetic Variants |
| title_full | Development of Conventional Multiplex PCR Assays for the Identification of 21 West Palaearctic Biting Midge Taxa (Diptera: Ceratopogonidae) Belonging to the <i>Culicoides</i> Subgenus <i>Culicoides</i>, including Recently Discovered Species and Genetic Variants |
| title_fullStr | Development of Conventional Multiplex PCR Assays for the Identification of 21 West Palaearctic Biting Midge Taxa (Diptera: Ceratopogonidae) Belonging to the <i>Culicoides</i> Subgenus <i>Culicoides</i>, including Recently Discovered Species and Genetic Variants |
| title_full_unstemmed | Development of Conventional Multiplex PCR Assays for the Identification of 21 West Palaearctic Biting Midge Taxa (Diptera: Ceratopogonidae) Belonging to the <i>Culicoides</i> Subgenus <i>Culicoides</i>, including Recently Discovered Species and Genetic Variants |
| title_short | Development of Conventional Multiplex PCR Assays for the Identification of 21 West Palaearctic Biting Midge Taxa (Diptera: Ceratopogonidae) Belonging to the <i>Culicoides</i> Subgenus <i>Culicoides</i>, including Recently Discovered Species and Genetic Variants |
| title_sort | development of conventional multiplex pcr assays for the identification of 21 west palaearctic biting midge taxa diptera ceratopogonidae belonging to the i culicoides i subgenus i culicoides i including recently discovered species and genetic variants |
| topic | <i>Culicoides</i> Pulicaris Group West Palaearctic vectors polymerase chain reaction (PCR) mitochondrial cytochrome c oxidase subunit I (COI) |
| url | https://www.mdpi.com/1424-2818/15/6/699 |
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