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Application of newer technologies for the diagnosis of FMD and other vesicular diseases - SE1120

Description
In the event of a suspected occurence of vesicular disease, rapid confirmation by laboratory investigation of index case is required. In the event of secondary outbreaks arising then the emphasis will shift to moving rapid diagnosis of cases onto farms (pen-side tests) to reduce or avoid delays associated with referral to reference laboratories. Such an approach would enable the clinical suspicion of disease to be quickly investigated and supported by objective analysis, to ensure that the presence of virus is quickly identified and speedily eliminated to control the spread of infection or to identify antibody from serological survey.

Diagnostic tests need to be highly sensitive, specific and swift. The current validated laboratory procedures for foot-and-mouth (FMD) and related vesicular virus diagnosis are antigenic detection ELISA's combined with virus isolation in sensitive cell cultures. Real time fluorogenic reverse transcripton polymerase chain reaction (RT-PCR) assays are likely to provide a marked increase in test sensitivity and faster definitive diagnostic results. However, the procedures require further development and validation and are the subject of a separate project proposal. The assay employs polyclonal antisera. The incorporation of monoclonal instead of polyclonal reagents will be addressed to overcome limitations of finite supply of individual polyclonal antiserum batches. This will improve specificity and standardisation and may improve sensitivity and speed. Recombinant FMDV receptor protins and mabs to additional vesicular viruses will also be evaluated as possible virus trapping ligands for use in antigen detection assays.

FMD mabs have already been incoprated into prototype chromatographic strip tests for pen-side antigen detection. These tests will be evaluated by means of field trials in countries with endemic FMD. Prototype pen-side tests for additional vesicular viruses will be developed. Other innovative diagnostic approaches will be expored, such as piezo-electric immuobiosensors and micro-arrays. The former has potential for penside application, and the latter for differential diagnosis.

Pan-reactive mabs able to detect all serotypes of FMDV willa lso be evaluated for possible used in serotype-independent competitive ELISA for the detection of antibodies. If promising such reagents could alo be applied to pen-side tests.
Objective
01: Evaluate FMD mabs and recombiant integrins for inter- and intra-typic reactivity for use in the indirect sandwich ELISA for antigen detection, in chromatographic strip test devices, in piezo-electric bisensors and in the competition ELISA for serology (23). Produce further mabs if gaps exist in the collection.

02: Develop prototype pen-side tests for FMD, SVD and VS diagnosis.

03: Field validate FMD and other vesicular disease pen-side strip test devices.

04: Further evaluation o new cell lines and integrin transfected cells for FMD and other vesicular virus diagnosis.

05: Develop prototype micro-array test for vesicular viruses.
Project Documents
• Final Report : Application of newer technologies for the diagnosis of FMD and other vesicular diseases   (4245k)
Time-Scale and Cost
From: 2003

To: 2006

Cost: £396,290
Contractor / Funded Organisations
Institute for Animal Health (BBSRC)
Keywords
Animal Diseases              
Animal Health              
Biotechnology              
Diagnosis              
Foot and Mouth              
GM Non-Food              
Plants and Animals              
Fields of Study
Animal Health