title: Detection of African horse sickness virus in Culicoides imicola pools using RT-qPCR.
abstract: African horse sickness (AHS) is an infectious, non-contagious arthropod-borne disease of equids, caused by the African horse sickness virus (AHSV), an orbivirus of the Reoviridae family. It is endemic in sub-Saharan Africa and thought to be the most lethal viral disease of horses. This study focused on detection of AHSV in Culicoides imicola (Diptera: Ceratopogonidae) pools by the application of a RT-qPCR. Midges were fed on AHSV-infected blood. A single blood-engorged female was allocated to pools of unfed nulliparous female midges. Pool sizes varied from 1 to 200. RNA was extracted and prepared for RT-qPCR. The virus was successfully detected and the optimal pool size for the limit of detection of the virus was determined at a range between 1 to 25. Results from this investigation highlight the need for a standardized protocol for AHSV investigation in Culicoides midges especially for comparison among different studies and for the determination of infection rate.
authors.affiliation: Unit for Environmental Sciences and Management, North-West University, Potchefstroom campus, Private Bag X6001, Potchefstroom 2520, South Africa.
authors.forename: Tania
authors.initials: T
authors.lastname: de Waal
authors.affiliation: Unit for Environmental Sciences and Management, North-West University, Potchefstroom campus, Private Bag X6001, Potchefstroom 2520, South Africa. Danica.LiebenbergWeyers@nwu.ac.za.|Faculty of Education Sciences, School for Natural Sciences and Technology for Education, North-West University, Potchefstroom campus, Private Bag X6001, Potchefstroom 2520, South Africa. Danica.LiebenbergWeyers@nwu.ac.za.
authors.forename: Danica
authors.initials: D
authors.lastname: Liebenberg
authors.affiliation: Parasites, Vectors and Vector Borne diseases, Agricultural Research Council- Onderstepoort Veterinary Institute, Private Bag X5, Onderstepoort 0110, South Africa.
authors.forename: Gert J
authors.initials: GJ
authors.lastname: Venter
authors.affiliation: Unit for Environmental Sciences and Management, North-West University, Potchefstroom campus, Private Bag X6001, Potchefstroom 2520, South Africa.
authors.forename: Charlotte Ms
authors.initials: CM
authors.lastname: Mienie
authors.affiliation: Unit for Environmental Sciences and Management, North-West University, Potchefstroom campus, Private Bag X6001, Potchefstroom 2520, South Africa.
authors.forename: Huib
authors.initials: H
authors.lastname: van Hamburg
mesh_terms: D000355:African Horse Sickness; D000356:African Horse Sickness Virus / Q000302:isolation & purification*; D000818:Animals; D002519:Ceratopogonidae / Q000821:virology*; D005260:Female; D006736:Horses; D007303:Insect Vectors / Q000821:virology*; D060888:Real-Time Polymerase Chain Reaction
keywords: Pool size; RT-qPCR sensitivity; limit of detection