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

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.

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Main Authors: Scheffer,Elisabeth G., Venter,Gert J., Joone,Christopher, Osterrieder,Nikolaus, Guthrie,Alan J.
Format: Digital revista
Language:English
Published: ARC-Onderstepoort Veterinary Institute - Agricultural Research Council 2011
Online Access:http://www.scielo.org.za/scielo.php?script=sci_arttext&pid=S0030-24652011000100016
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spelling oai:scielo:S0030-246520110001000162012-07-27Use of real-time quantitative reverse transcription polymerase chain reaction for the detection of African horse sickness virus replication in Culicoides imicolaScheffer,Elisabeth G.Venter,Gert J.Joone,ChristopherOsterrieder,NikolausGuthrie,Alan J.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.ARC-Onderstepoort Veterinary Institute - Agricultural Research CouncilOnderstepoort Journal of Veterinary Research v.78 n.1 20112011-01-01journal articletext/htmlhttp://www.scielo.org.za/scielo.php?script=sci_arttext&pid=S0030-24652011000100016en
institution SCIELO
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country Sudáfrica
countrycode ZA
component Revista
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databasecode rev-scielo-za
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libraryname SciELO
language English
format Digital
author Scheffer,Elisabeth G.
Venter,Gert J.
Joone,Christopher
Osterrieder,Nikolaus
Guthrie,Alan J.
spellingShingle Scheffer,Elisabeth G.
Venter,Gert J.
Joone,Christopher
Osterrieder,Nikolaus
Guthrie,Alan J.
Use of real-time quantitative reverse transcription polymerase chain reaction for the detection of African horse sickness virus replication in Culicoides imicola
author_facet Scheffer,Elisabeth G.
Venter,Gert J.
Joone,Christopher
Osterrieder,Nikolaus
Guthrie,Alan J.
author_sort Scheffer,Elisabeth G.
title Use of real-time quantitative reverse transcription polymerase chain reaction for the detection of African horse sickness virus replication in Culicoides imicola
title_short Use of real-time quantitative reverse transcription polymerase chain reaction for the detection of African horse sickness virus replication in Culicoides imicola
title_full Use of real-time quantitative reverse transcription polymerase chain reaction for the detection of African horse sickness virus replication in Culicoides imicola
title_fullStr Use of real-time quantitative reverse transcription polymerase chain reaction for the detection of African horse sickness virus replication in Culicoides imicola
title_full_unstemmed Use of real-time quantitative reverse transcription polymerase chain reaction for the detection of African horse sickness virus replication in Culicoides imicola
title_sort use of real-time quantitative reverse transcription polymerase chain reaction for the detection of african horse sickness virus replication in culicoides imicola
description 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.
publisher ARC-Onderstepoort Veterinary Institute - Agricultural Research Council
publishDate 2011
url http://www.scielo.org.za/scielo.php?script=sci_arttext&pid=S0030-24652011000100016
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