Implicating Culicoides biting midges as vectors of Schmallenberg virus using semi-quantitative RT-PCR

The recent unprecedented emergence of arboviruses transmitted by Culicoides biting midges in northern Europe has necessitated the development of techniques to differentiate competent vector species. At present these techniques are entirely reliant upon interpretation of semi-quantitative RT-PCR (sqP...

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Bibliographic Details
Published in:PloS one 2013-03, Vol.8 (3), p.e57747-e57747
Main Authors: Veronesi, Eva, Henstock, Mark, Gubbins, Simon, Batten, Carrie, Manley, Robyn, Barber, James, Hoffmann, Bernd, Beer, Martin, Attoui, Houssam, Mertens, Peter Paul Clement, Carpenter, Simon
Format: Article
Language:English
Subjects:
Animals
Biology
Biting
Bunyaviridae Infections - genetics
Bunyaviridae Infections - transmission
Bunyaviridae Infections - virology
Ceratopogonidae
Ceratopogonidae - genetics
Ceratopogonidae - virology
Culicoides
Culicoides variipennis
Diagnostic systems
Diptera
Disease transmission
Genetic vectors
Infection
Infections
Insect Vectors - genetics
Insect Vectors - virology
Laboratories
Life Sciences
Mathematics
Medical research
Medicine
Orthobunyavirus - genetics
Polymerase chain reaction
Reverse Transcriptase Polymerase Chain Reaction - methods
Ribonucleic acid
RNA
Studies
Vectors
Vectors (Biology)
Veterinary Science
Virology
Viruses
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Summary:The recent unprecedented emergence of arboviruses transmitted by Culicoides biting midges in northern Europe has necessitated the development of techniques to differentiate competent vector species. At present these techniques are entirely reliant upon interpretation of semi-quantitative RT-PCR (sqPCR) data in the form of Cq values used to infer the presence of viral RNA in samples. This study investigates the advantages and limitations of sqPCR in this role by comparing infection and dissemination rates of Schmallenberg virus (SBV) in two colony lines of Culicoides. Through the use of these behaviorally malleable lines we provide tools for demarcating arbovirus infection and dissemination rates in Culicoides which to date have prevented clear implication of primary vector species in northern Europe. The study demonstrates biological transmission of SBV in an arthropod vector, supporting the conclusions from field-caught Culicoides and provides a general framework for future assessment of vector competence of Culicoides for arboviruses using sqPCR. When adopting novel diagnostic technologies, correctly implicating vectors of arboviral pathogens requires a coherent laboratory framework to fully understand the implications of results produced in the field. This study illustrates these difficulties and provides a full examination of sqPCR in this role for the Culicoides-arbovirus system.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0057747