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Epidemic hemorrhagic disease virus (EHDV) is a member of the genus Orbivirus of the family Reoviridae, which has a double-layered capsid structure. The outer capsid surface is composed of 42 ring-shaped capsid particles, which are typical features of the virus. The genome of EHDV is a double-stranded RNA of about 20 kb, which can encode 7 structural proteins (VP1-VP7) and 4 non-structural proteins (NS1, NS2, NS3, NS3a). Epidemic hemorrhagic disease (EHD) is an arboviral disease caused by EHDV infection of ruminants. EHD is prevalent in tropical, subtropical and temperate regions and is transmitted by the blood-sucking bite of female Culicoides. EHD seriously affects the development of animal husbandry and international trade in various countries, and has been classified as a legally reported infectious disease by the World Organization for Animal Health (OIE).
Figure 1. Life cycle of epizootic hemorrhagic disease and bluetongue (Rivera, et al. 2021).
Epizootic Hemorrhagic Disease Virus PCR Kit Development
The VP2 protein encoded by Seg-2 is the main protein that constitutes the outer coat of EHDV and is highly variable. It is the main protein that induces the production of specific neutralizing antibodies and determines the serotype of the virus. At present, at least 9 different serotypes of EHDV have been identified worldwide, and the RNA fragment 2 encoding VP2 is the target of PCR serotyping. The researchers designed specific primer sequences according to Seg-2 of EHDV of different serotypes, and developed a variety of PCR detection methods, including RT-PCR, qRT-PCR, multiplex RT-PCR, real-time RT-PCR, etc. The development of these molecular diagnostic methods provides the basis for the design and timely implementation of EHD control measures.
BioVenic is a provider of biology reagents and kits, which has been committed to the development of animal pathogens diagnostic reagents for many years. We have a professional team with extensive experience in veterinary diagnostic and reagent development, and our R&D team is working hard to develop and optimize PCR kits. We can provide you with a series of customized PCR kits for the detection of epidemic hemorrhagic disease virus according to your needs. If you have any needs, please feel free to contact us. We will provide you with high-quality products and services.
Epizootic Hemorrhagic Disease Virus PCR Kit We Can Develop
| EHDV RT-PCR kit | EHDV nested PCR kit | EHDV real-time PCR kit |
| EHDV RT-qPCR kit | EHDV multiplex RT-PCR Kit | Other EHDV PCR Kit |
Workflow of Epizootic Hemorrhagic Disease Virus PCR Kit Development
| Workflow | Details |
| Samples and cell cultures | Samples were obtained from naturally infected cattle or buffalo, or from experimentally infected animals. EHDV strains were grown in bovine cells. |
| Primer and probes design | Design primers or probes to detect all serotypes and fragment 2 for serotyping. Use Primer Express software to select and evaluate primers and probes. |
| Nucleic acid extraction | Extract RNA from blood of infected animals. |
| Establishment of PCR conditions | Establishment of thermocycling conditions to conduct amplification and detection through PCR. |
| Reproducibility evaluation of the PCR assay | The reproducibility of the PCR kit was assessed by determining the intra-assay and inter-assay coefficients of variation (CV). |
| Assessment of sensitivity and specificity | A panel of different EHDV strains, including all serotypes, was tested to assess the specificity of PCR kits. In addition, the serotypes of other pathogens such as bluetongue virus are also tested to ensure that there is no cross-reactivity. Determine the minimum amount of EHDV detected in the sample to assess the sensitivity of the PCR kit. |
Delivery
- A series of PCR kits for epidemic hemorrhagic disease virus
- Product quality inspection report
- Other experimental data you need
Our Advantages
- High specificity and sensitivity
- High reproducibility between tests
- Detects multiple serological EHDVs
- Faster results and greater cost savings
References
- Rivera, Nelda A., et al. "Bluetongue and epizootic hemorrhagic disease in the United States of America at the wildlife–livestock interface." Pathogens 10.8 (2021): 915.
- Clavijo, Alfonso, et al. "An improved real-time polymerase chain reaction for the simultaneous detection of all serotypes of Epizootic hemorrhagic disease virus." Journal of Veterinary Diagnostic Investigation 22.4 (2010): 588-593.
- Viarouge, Cyril, et al. "Duplex real-time RT-PCR assays for the detection and typing of epizootic haemorrhagic disease virus." PLoS One 10.7 (2015): e0132540.
