Real-time fluorogenic reverse transcription polymerase chain reaction assay for detection of African horse sickness virus.
Abstract: African horse sickness is an arthropod-borne disease of the equine included in the World Organization for Animal Health (OIE) list with important economic consequences for horse trade. The disease is caused by African horse sickness virus (AHSV; family Reoviridae, genus Orbivirus), which is transmitted by Culicoides midges. It is endemic in sub-Saharan Africa, spreading occasionally outside this area where the occurrence of Culicoides vectors allows virus transmission. Currently, only conventional (gel-based) reverse transcription polymerase chain reaction (RT-PCR) protocols are available for its detection; however, these methods are cumbersome and difficult to apply when large numbers of samples are to be tested, as in the case of epizootics. To overcome this problem, a real-time RT-PCR method has been developed, based on a 5'-Taq nuclease-3'-minor groove binder-DNA probe (TaqMan MGB) for detection of a wide range of AHSV serotypes and strains designed to the highly conserved region of the VP7 gene (segment 7). The method was able to detect all prototype strains from the 9 known serotypes of the virus, with a high analytical sensitivity; no cross-reactions were observed with other orbiviruses or with other viruses affecting horses. The diagnostic sensitivity was assessed using a panel of AHSV-positive tissue samples from an epizootic that occurred in Spain between 1987 and 1990. This method, which can be performed in 96-well format, is suitable for large-scale surveillance of AHSV in areas where it can potentially spread.
Publication Date: 2008-05-08 PubMed ID: 18460619DOI: 10.1177/104063870802000310Google Scholar: Lookup
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- Journal Article
- African Horse Sickness
- Culicoides
- Diagnosis
- Diagnostic Technique
- Disease Diagnosis
- Disease Outbreaks
- Disease Surveillance
- Economics
- Epidemiology
- Epizootic
- Equine Diseases
- Equine Health
- Infectious Disease
- Laboratory Methods
- Polymerase Chain Reaction
- Real-Time PCR
- Serotypes
- Vector-borne disease
- Veterinary Medicine
- Virology
- Virus
Summary
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The research article discusses the development of a more efficient detection method for African horse sickness virus (AHSV), a serious disease affecting horses with high economic impact. The newly proposed method aims to overcome the limitations of existing detection protocols, leveraging real-time RT-PCR technology for better accuracy and feasibility in large-scale testing scenarios.
Understanding the Context: African Horse Sickness and Its Impact
- African horse sickness (AHS) is a disease that affects equines and is significant enough to be featured on the World Organization for Animal Health’s list.
- The disease is caused by the African horse sickness virus (AHSV), a type of Orbivirus from the Reoviridae family, and is transmitted by Culicoides midges.
- While AHSV is endemic in sub-Saharan Africa, it occasionally spreads outside this region, making it an area of significant concern for horse health and global trade.
The Current Detection Method and its Limitations
- The existing method for detecting AHSV is through conventional reverse transcription polymerase chain reaction (RT-PCR) protocols.
- While effective, these methods are intricate and not conducive to large-scale testing, such as during large disease outbreaks.
A New Approach: Real-Time RT-PCR
- The researchers developed a new detection method, a real-time RT-PCR method, to overcome the limitations of the conventional approach.
- This improved method uses a 5′-Taq nuclease-3′-minor groove binder-DNA probe, designed to target the highly conserved region of the VP7 gene, an integral component of the AHSV.
- The real-time RT-PCR method was successful in detecting all prototype strains from all known serotypes of the virus, demonstrating high analytical sensitivity.
- The method showed no cross-reactions with other orbiviruses or with other viruses affecting horses, implying its specificity to AHSV.
Validation and Potential Use
- The diagnostic sensitivity of the new method was verified using a panel of AHSV-positive tissue samples from an outbreak that took place in Spain between 1987 and 1990.
- This innovative method, which can be executed in a 96-well format, is ideal for large-scale AHSV surveillance, especially in areas where the disease could potentially spread.
The results of the research highlight a significant advancement in detecting the AHSV, which could have profound consequences for our ability to control and manage the spread of African horse sickness in different regions around the world.
Cite This Article
APA
Agüero M, Gómez-Tejedor C, Angeles Cubillo M, Rubio C, Romero E, Jiménez-Clavero A.
(2008).
Real-time fluorogenic reverse transcription polymerase chain reaction assay for detection of African horse sickness virus.
J Vet Diagn Invest, 20(3), 325-328.
https://doi.org/10.1177/104063870802000310 Publication
Researcher Affiliations
- Laboratorio Central de Veterinaria, Algete, Madrid, Spain.
MeSH Terms
- African Horse Sickness / diagnosis
- African Horse Sickness / virology
- African Horse Sickness Virus
- Animals
- Horses
- Reverse Transcriptase Polymerase Chain Reaction / methods
- Reverse Transcriptase Polymerase Chain Reaction / veterinary
- Sensitivity and Specificity
- Spleen / virology
Citations
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