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Molecular detection techniques to investigate aspects of the pathogenesis of rabies - SE0413

Description
The primary objective of this research is to investigate the pathogenesis of disease using the group's recently developed, highly sensitive Polymerase Chain Reaction (PCR)- based techniques for the detection of rabies virus (RV) and rabies-related virus (RRV) in a variety of tissues. This PCR-based test system will be further developed in order to:

* Monitor the excretion of RV in the saliva of foxes and dogs to elucidate patterns of virus excretion over the critical pre-clinical and prodromal stages of disease.
* Detect virus in RV and European Bat Lyssaviruses (EBL)- infected mice throughout the post-infection disease process to clarify further the pathogenesis of RV and EBL infection.
* Detect virus in tissue and blood samples from vaccinated mice which have been challenged with RV and RRVs

In addition, the antibody response of infection and vaccinated mice which have been challenged with RV and RRVs.

In additon, the antibody response of infected and vaccinated mice will be monitored by the conventional test methodology and Western blot assays (WBAs). The aim of these studies will be to elucidate the humoral immune response to RV and the EBLs and to investigate the extent to which the currently avaliable HDC vaccine confers protection against the EBLs.

This research will enhance existing knowledge of the pathogenesis and humoral immune response involved in RV and RRV infection. A primary aim of this research is to develop a test able to confirm whether an animal is excreting RV in its saliva and therefore able to identify those animals able to transmit rabies. This test would underpin the early decision-making involved in implementing contingency plans in the event of a rabies outbreak in the UK. In addition, it is envisaged that enhanced tests for use in ante-mortem and routine rabies diagnosis will arise from these studies. This research will fulfil a ROAME A objective to investigate the pathogenesis of RV infection using a sensitive molecular test system.
Objective
01 01/04/99 Develop a PCR- based test fro the detection of Rv in salvia

02 31/05/00 Monitor RV levels in the saliva of infected foxes and dogs.

03 31/03/02 Develop a Western blot assay for assessment of antibody response to Rv, RRVs and vaccine strains.

04 31/03/02 Monitor virus presence in mice infected with RV and EBLs 1 and 2 during the post-infection phases by testing RNA from slected tissues at vairous times post-infection using hnRT-PCR.

05 31/03/02 Detect virus in vaccinated mice which have been subsquently challenged with RV and EBLs 1 and 2 by testing RNA from selected tissues using hnRT-PCR.
Project Documents
• Final Report : Molecular Detection Techniques to Investigate Aspects of the Pathogenesis of Rabies   (76k)
Time-Scale and Cost
From: 1998

To: 2002

Cost: £615,773
Contractor / Funded Organisations
Veterinary Laboratories Agency
Keywords
Animal Health              
Biotechnology              
Diagnosis              
Pathogenesis              
Plants and Animals              
Rabies              
Zoonoses              
Fields of Study
Animal Health