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Application of newer technologies for the diagnosis of FMD and other vesicular diseases - SE1117

Description

In the event of an outbreak of vesicular disease rapid confirmation by laboratory investigation is required. The current validated procedures for foot-and-mouth disease (FMD) and related vesicular virus diagnosis are antigen detection ELISA’s combined with virus isolation in sensitive cell cultures. Research is aimed at improving the sensitivity and specificity of diagnostic tests for vesicular virus antigen detection. The current ELISA for the antigen detection of FMD virus is highly sensitive but employs polyclonal antisera. The incorporation of monoclonal instead of polyclonal reagents would overcome limitations of finite supply of individual polyclonal antiserum batches and should improve specificity and standardisation. The FMD monoclonal antibodies will also be investigated for likely incorporation into a competitive ELISA for the detection of antibodies and into chromatographic strip-test devices for pen-side diagnosis. Calf thyroid (CTY) cells are the most sensitive cell culture detection system for the majority of wild-type FMD virus. However, virus sensitivity of these cultures is rapidly lost upon passage and thyroid glands have to be regularly obtained to yield primary cell cultures. Attempts will be made to create stable cell lines transfected with the FMD virus integrin receptor as a means of improving their sensitivity to infection and to provide an alternative system to primary CTY cells for FMD virus diagnosis. Rapid, on farm diagnosis will be required in the future to quickly investigate the clinical suspicion of disease to ensure that the presence of virus is quickly identified and speedily eliminated or to identify antibody from serological survey. One approach is through the use of chromatographic strip test devices incorporating vesicular disease virus mabs or recombinant antigens as ligands. These will be devised and evaluated for the detection of vesicular disease virus antigens and antibody and for their possible application to pen-side diagnosis.

Objective

The project has three main objectives :

1. Produce and evaluate monoclonal antibody (mabs) to FMD virus with broad intra-typic reactivity for use in the indirect, sandwich ELISA for antigen detection, in chromatographic strip test devices and in the competition ELISA for serology (18).

2. Identify or establish immortalised cell lines as alternative cell culture systems to primary CTY cells for FMD virus isolation and ascertain their sensitivity to FMD (and other vesicular) virus growth.

3. Produce and evaluate rapid chromatographic strip-test devices for pen-side vesicular virus disease diagnosis.

Project Documents
• Final Report : SE1117 Application of newer technologies for the diagnosis of FMD and other vesicular diseases   (183k)
Time-Scale and Cost
From: 2000

To: 2003

Cost: £315,918
Contractor / Funded Organisations
Institute for Animal Health (BBSRC)
Keywords
Animal Diseases              
Animal Health              
Biotechnology              
Control              
Diagnosis              
Foot and Mouth              
Plants and Animals              
Fields of Study
Animal Health