Original Research

Use of real-time quantitative reverse transcription polymerase chain reaction for the detection of African horse sickness virus replication in Culicoides imicola

Elisabeth G. Scheffer, Gert J. Venter, Christopher Joone, Nikolaus Osterrieder, Alan J. Guthrie
Onderstepoort Journal of Veterinary Research | Vol 78, No 1 | a344 | DOI: https://doi.org/10.4102/ojvr.v78i1.344 | © 2011 Elisabeth G. Scheffer, Gert J. Venter, Christopher Joone, Nikolaus Osterrieder, Alan J. Guthrie | This work is licensed under CC Attribution-NoDerivatives 4.0
Submitted: 21 May 2011 | Published: 11 November 2011

About the author(s)

Elisabeth G. Scheffer, Equine Research Centre, University of Pretoria, South Africa
Gert J. Venter, Parasites, Vectors and Vector-borne Diseases, ARC-Onderstepoort Veterinary Institute, South Africa
Christopher Joone, Equine Research Centre, University of Pretoria, South Africa
Nikolaus Osterrieder, Institut für Virologie, Freie Universität Berlin, Germany, Germany
Alan J. Guthrie, Equine Research Centre, University of Pretoria, South Africa


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Abstract

Despite its important role as vector for African horse sickness virus (AHSV), very little information is available on the dissemination of this virus in Culicoides (Avaritia) imicola Kieffer (Diptera: Ceratopogonidae). This study reports on the applicability of a real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) to detect AHSV in dissected midges. A total of 96 midges were fed on AHSV-infected blood, after which one test group was dissected into head/thorax and abdomen segments immediately after feeding and the other only after 10 days of incubation. The majority of the midges (96%) ingested the virus successfully and there was no significant difference between the virus concentration in the heads/thoraxes and the abdomens immediately after feeding. After incubation, virus was detected in 51% of the midges and it was confined to the abdomen in the majority of these. The fact that virus was detected only in the heads/thoraxes of four Culicoides midges after incubation suggests the presence of a mesenteronal escape barrier. Replication in the salivary glands was not shown. An increase of the mean virus concentration in the abdomen after incubation indicates localised viral replication. The real-time RT-qPCR is recommended for further studies investigating the replication and dissemination of AHSV in Culicoides midges.

Keywords

African horse sickness virus; Culicoides; polymerase chain reaction; vector; virus replication

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Crossref Citations

1. Detection of African horse sickness virus inCulicoides imicolapools using RT-qPCR
Tania de Waal, Danica Liebenberg, Gert J Venter, Charlotte MS Mienie, Huib van Hamburg
Journal of Vector Ecology  vol: 41  issue: 1  first page: 179  year: 2016  
doi: 10.1111/jvec.12210