Header

UZH-Logo

Maintenance Infos

Molecular characterisation of Swiss Ceratopogonidae (Diptera) and evaluation of real-time PCR assays for the identification of Culicoides biting midges


Wenk, Claudia E. Molecular characterisation of Swiss Ceratopogonidae (Diptera) and evaluation of real-time PCR assays for the identification of Culicoides biting midges. 2011, University of Zurich, Vetsuisse Faculty.

Abstract

Biting midges of the genus Culicoides (Diptera, Ceratopogonidae) are vectors of several viruses of veterinary relevance, and they can cause insect bite hypersensitivity. As the morphological identification of these tiny insects is difficult, we aimed at developing real-time PCRs. Partial mitochondrial cytochrome oxidase I gene (mtCOI) sequences were determined from 380 Culicoides midges from three regions of Switzerland (Alps, Midland, Ticino) and from non-biting midges. Sequence variability (haplotypes) was observed in all 21 morphologically identified species. For each of C. grisescens and C. obsoletus, a cryptic species was identified. Further, a sister taxon to C. pulicaris was determined (Culicoides sp.). Alignments of available mtCOI sequences from Ceratopogonidae (GenBank, this study) were used to design real-time PCRs to distinguish C. chiopterus, C. deltus, C. dewulfi, C. grisescens, C. imicola, C. lupicaris, C. obsoletus (both species), C. pulicaris, C. scoticus and Culicoides sp. Specificities of the assays were tested with cloned targets from 1 to 4 haplotypes of 18 Culicoides spp. and 4 other Ceratopogonidae revealing no cross-reactivity when testing 5x106 gene copies. The sensitivities of two assays were tested by spiking single insects into pools of 99 or 999 randomly selected non-target Ceratopogonidae. Primers and probes of this study were devised to be suitable for multiplexed assays but these evaluations await to be performed.

Abstract

Biting midges of the genus Culicoides (Diptera, Ceratopogonidae) are vectors of several viruses of veterinary relevance, and they can cause insect bite hypersensitivity. As the morphological identification of these tiny insects is difficult, we aimed at developing real-time PCRs. Partial mitochondrial cytochrome oxidase I gene (mtCOI) sequences were determined from 380 Culicoides midges from three regions of Switzerland (Alps, Midland, Ticino) and from non-biting midges. Sequence variability (haplotypes) was observed in all 21 morphologically identified species. For each of C. grisescens and C. obsoletus, a cryptic species was identified. Further, a sister taxon to C. pulicaris was determined (Culicoides sp.). Alignments of available mtCOI sequences from Ceratopogonidae (GenBank, this study) were used to design real-time PCRs to distinguish C. chiopterus, C. deltus, C. dewulfi, C. grisescens, C. imicola, C. lupicaris, C. obsoletus (both species), C. pulicaris, C. scoticus and Culicoides sp. Specificities of the assays were tested with cloned targets from 1 to 4 haplotypes of 18 Culicoides spp. and 4 other Ceratopogonidae revealing no cross-reactivity when testing 5x106 gene copies. The sensitivities of two assays were tested by spiking single insects into pools of 99 or 999 randomly selected non-target Ceratopogonidae. Primers and probes of this study were devised to be suitable for multiplexed assays but these evaluations await to be performed.

Statistics

Downloads

10 downloads since deposited on 29 Apr 2019
10 downloads since 12 months
Detailed statistics

Additional indexing

Other titles:Molekulare Charakterisierung von Ceratopogoniden aus der Schweiz und Evaluation von real-time PCR Assays zur Identifizierung von Culicoi- des Gnitzen
Item Type:Dissertation (monographical)
Referees:Mathis Alexander, Engels Monika
Communities & Collections:05 Vetsuisse Faculty > Institute of Parasitology
04 Faculty of Medicine > Institute of Parasitology

UZH Dissertations
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
600 Technology
Uncontrolled Keywords:Culicoides, vector identification, barcode, real-time PCR, specificity, cryptic species
Language:English
Place of Publication:Zürich
Date:2011
Deposited On:29 Apr 2019 16:16
Last Modified:03 Oct 2019 08:24
Number of Pages:12
Additional Information:Publiziert in und Sonderdruck aus: Veterinary Parasitology, 184 (2012), 258-266
OA Status:Green
Related URLs:https://www.recherche-portal.ch/primo-explore/fulldisplay?docid=ebi01_prod007015077&context=L&vid=ZAD&search_scope=default_scope&tab=default_tab&lang=de_DE (Library Catalogue)

Download

Green Open Access

Download PDF  'Molecular characterisation of Swiss Ceratopogonidae (Diptera) and evaluation of real-time PCR assays for the identification of Culicoides biting midges'.
Preview
Content: Published Version
Language: English
Filetype: PDF
Size: 571kB