Epizootic Hemorrhagic Disease Virus Methods and Protocols

This volume provides detailed methods to support fast and reliable diagnosis, surveillance and control of Epizootic Haemorrhagic disease virus (EHDV), with the aim of facilitating and promoting further research into this understudied Orbivirus. Chapters guide readers through methods to enable the characterization of EHDV in vitro and in vivo. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Epizootic Hemorrhagic Disease Virus: Methods and Protocols aims to be a useful up-to-date reference work for researchers currently in the field and to those entering the field.

Managing risk when working with Orbiviruses with a special emphasis on Epizootic haemorrhagic disease virus.- Recognition of Field Signs, Necropsy Procedures and Evaluation of Macroscopic Lesions of Cervids Infected with Epizootic Hemorrhagic Disease Virus.- Virus Isolation of Epizootic Haemorrhagic Disease Virus in Cell Culture.- Purification of epizootic haemorrhagic disease virus (and other Orbiviruses) particles from infected mammalian or insect cells.- Confocal microscopy of fluorescently labelled EHDV infected cell cultures.- Epizootic Haemorrhagic Disease Virus Titration.- Virus neutralisation test for detecting and quantifying serum neutralising antibodies to epizootic haemorrhagic disease virus (serotypes 1, 2, and 4-8).- Epizootic Hemorrhagic Disease Agar Gel Immunodiffusion Assay.- Detection of EHDV antibodies using enzyme-linked immunosorbent assay (ELISA).- Real-time RT-PCR for the diagnosis of epizootic haemorrhagic disease virus.- Real-time RT-PCR assays for typing of Epizootic Haemorrhagic Disease virus.- Rapid Laboratory Diagnosis of Epizootic Haemorrhagic Disease Virus using a Loop Mediated Isothermal Amplification Assay.- End-point RT-PCR assays for detection and typing of Epizootic Haemorrhagic Disease virus.- Molecular serotyping of epizootic haemorrhagic disease virus.-Whole genome sequencing of Epizootic Haemorrhagic Disease Virus.- Infection of Culicoides biting midges with Epizootic Haemorrhagic Disease Virus.