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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.
© 2016 The Society for Vector Ecology.
Publication Date: 2016-05-28 PubMed ID: 27232141DOI: 10.1111/jvec.12210Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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The research demonstrates a method to detect African horse sickness virus in specific types of midges using reverse transcription quantitative PCR. This study also identified the optimal sample pool size for accurate detection.
Understanding the Disease and Its Carrier
- African horse sickness (AHS) is a dangerous viral disease that affects horses and other equids. It is most common in sub-Saharan Africa and is known for its deadly effects. The virus causing AHS is carried by a specific type of midge, called Culicoides imicola.
Methodology
- The researchers fed the midges on blood that was infected with the African horse sickness virus (AHSV).
- After the feeding, a single infected midge was added to groups (pools) of other unfed midges. These pool sizes ranged from 1 to 200 midges.
- Then, the researchers extracted RNA from these pools and prepared it for reverse transcription quantitative PCR (RT-qPCR), which is a testing method used to detect specific sequences of DNA or RNA in a sample.
Results and Conclusions
- The RT-qPCR method successfully detected the presence of AHSV in the midges.
- The study also determined that pool sizes between 1 and 25 were most effective for accurately detecting the virus.
- The findings highlight the importance of creating a standard procedure for detecting AHSV in Culicoides midges to ensure accurate comparison across various studies and to determine infection rates.
Implications of the study
- The conclusions drawn from this study can aid in better understanding of the spread of AHS and for developing future strategies for controlling the disease.
- The research can also help other studies that rely on the detection of AHSV in Culicoides midges to ensure they get accurate and comparable results.
Cite This Article
APA
de Waal T, Liebenberg D, Venter GJ, Mienie CM, van Hamburg H.
(2016).
Detection of African horse sickness virus in Culicoides imicola pools using RT-qPCR.
J Vector Ecol, 41(1), 179-185.
https://doi.org/10.1111/jvec.12210 Publication
Researcher Affiliations
- Unit for Environmental Sciences and Management, North-West University, Potchefstroom campus, Private Bag X6001, Potchefstroom 2520, South Africa.
- 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.
- Parasites, Vectors and Vector Borne diseases, Agricultural Research Council- Onderstepoort Veterinary Institute, Private Bag X5, Onderstepoort 0110, South Africa.
- Unit for Environmental Sciences and Management, North-West University, Potchefstroom campus, Private Bag X6001, Potchefstroom 2520, South Africa.
- Unit for Environmental Sciences and Management, North-West University, Potchefstroom campus, Private Bag X6001, Potchefstroom 2520, South Africa.
MeSH Terms
- African Horse Sickness
- African Horse Sickness Virus / isolation & purification
- Animals
- Ceratopogonidae / virology
- Female
- Horses
- Insect Vectors / virology
- Real-Time Polymerase Chain Reaction
Citations
This article has been cited 9 times.- Zhang Y, Zheng J, Zhang H, Lin Y, Wang Y, Ma Z, Wei J, Zhou B, Zhong D. Molecular and Serological Surveillance of Mosquito-Borne Viruses in Racehorses or Mosquitoes From Horse Farms in Shanghai, China, 2022. Transbound Emerg Dis 2025;2025:6131435.
- Li N, Meng J, He Y, Wang W, Wang J. Potential roles of Culicoides spp. (Culicoides imicola, Culicoides oxystoma) as biological vectors of bluetongue virus in Yuanyang of Yunnan, P. R. China. Front Cell Infect Microbiol 2023;13:1283216.
- Kampen H, Werner D. Biting Midges (Diptera: Ceratopogonidae) as Vectors of Viruses. Microorganisms 2023 Nov 4;11(11).
- Duan YL, Yang ZX, Bellis G, Li L. Isolation of Tibet Orbivirus from Culicoides jacobsoni (Diptera, Ceratopogonidae) in China. Parasit Vectors 2021 Aug 28;14(1):432.
- Barnes AN, Davaasuren A, Baasandavga U, Lantos PM, Gonchigoo B, Gray GC. Zoonotic enteric parasites in Mongolian people, animals, and the environment: Using One Health to address shared pathogens. PLoS Negl Trop Dis 2021 Jul;15(7):e0009543.
- Raksakoon C, Potiwat R. Current Arboviral Threats and Their Potential Vectors in Thailand. Pathogens 2021 Jan 18;10(1).
- Leta S, Fetene E, Mulatu T, Amenu K, Jaleta MB, Beyene TJ, Negussie H, Revie CW. Modeling the global distribution of Culicoides imicola: an Ensemble approach. Sci Rep 2019 Oct 2;9(1):14187.
- Leta S, Fetene E, Mulatu T, Amenu K, Jaleta MB, Beyene TJ, Negussie H, Kriticos D, Revie CW. Updating the global occurrence of Culicoides imicola, a vector for emerging viral diseases. Sci Data 2019 Sep 30;6(1):185.
- Wakefield A, Broyles M, Stone EL, Jones G, Harris S. Experimentally comparing the attractiveness of domestic lights to insects: Do LEDs attract fewer insects than conventional light types?. Ecol Evol 2016 Nov;6(22):8028-8036.
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