Scheffer, Elizabeth G.Venter, Gert JohannesJoone, ChristopherOsterrieder, NikolausGuthrie, Alan John2012-03-052012-03-052011-11-11Scheffer, E.G., Venter, G.J., Joone, C., Osterrieder, N. & Guthrie, A.J., 2011, ‘Use of real-time quantitative reverse transcription polymerase chain reaction for the detection of African horse sickness virus replication in Culicoides imicola’, Onderstepoort Journal of Veterinary Research 78(1), Art. #344, 4 pages. DOI: 10.4102/ojvr.v78i1.3440030-246510.4102/ojvr.v78i1.3447006535470J-6375-20130000-0001-7729-9918http://hdl.handle.net/2263/18374Despite 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.en© 2011. The Authors. Licensee: AOSIS OpenJournals. This work is licensed under the Creative Commons Attribution License.Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR)African horse sickness virusPolymerase chain reactionCulicoidesArticle