Abstract

Epizootie hemorrhagic disease of deer virus (EHDV), is a member of the family Reoviridae virus (Orbi virus), and a double-stranded RNA virus from arthropods (mainly Culicoides), which is very similar in morphology and structure to Blue tongue virus (BTV). Serologically, there is a clear cross between the blue tongue virus and EHDV. EHD (caused by EHDV) is a seasonal, non-contact viral infectious disease. Under natural conditions, EHDV can cause infection of cattle, sheep and other domestic animals as well as many domesticated and wild ruminants such as white tail deer, Mi, large horned antelope and so on. The virus has been isolated from white-tailed deer, black-tailed deer, sheep, goats, cattle, bison and Culicoides. White tail deer infection is the most serious, and clinical characteristics are transient elevated body temperature, symptoms of shock, mucosal, serous bleeding and death in a coma [1]. At present, there is no effective vaccine and treatment for this disease. To control and eliminate EHD, it is necessary to spend a lot of manpower and material resources. Export trade is restricted, which brings great economic losses to the countries with EHD epidemic. Moreover, because of the complexity of antigen structure and the diversity of clinical symptoms of EHDV, it is difficult to diagnose in laboratory. Therefore, developing a quick and precise laboratory diagnostic method becomes more and more significant among researchers.

Highlights

  • Epizootie hemorrhagic disease of deer virus (EHDV), is a member of the family Reoviridae virus (Orbi virus), and a double-stranded RNA virus from arthropods, which is very similar in morphology and structure to Blue tongue virus (BTV)

  • There is a clear cross between the blue tongue virus and EHDV

  • Because of the complexity of antigen structure and the diversity of clinical symptoms of EHDV, it is difficult to diagnose in laboratory

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Summary

Reagent

DEPC water, 0.01 mol/L PBS, Trizol lysate, trichloromethane, 75% ethyl alcohol, isopropyl alcohol (−20 ̊C precooling), other reagents are analytical pure, and grade-1 water as required by GB/T 6682. All reagents are separated by containers without RNA. By consulting the data and nucleic acid sequence of GenBank, the specific primers and probes were designed and synthesized for the gene sequence of EHDV: Forward primer 5’-GCGTTGGATATATTGGACAAAGC-3’; Reverse primer 5’-GCATACGAAGCATAAGCAACCTT-3’. TaqMan probe FAM-TCAAATCAAACGGGCGCAACTATGG-BGH, made into 5 umol/L, −20 ̊C. All are synthesized by Bao biologic engineering (Dalian) Co., Ltd

Sample Collection and Processing
Real-Time Fluorescent PCR Reaction
Methods
Building of Fluorescent Quantitative PCR Method
Specificity Test
Sensitivity and Reproducibility Test
Conclusion
Full Text
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