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Application of RT-PCR for the diagnosis of FMD and other vesicular diseases 1 - SE1119

In the event of an outbreak of vesicular disease rapid confirmation by laboratory investigation is required. Research is aimed at improving the sensitivity and specificity of diagnostic tests for detection of foot-and-mouth disease (FMD) and related vesicular viruses, as well as tyhe throughput and the speed of issue of final definitive resulta. Current procedures for vesicular virus investigation rely upon the use of ELISA for antigen detection coupled with virus isolation in cell cultrue. The former procedure is specific butnot sufficiently sensitive to detect 100% of positive samples thus necessitating the passage of the sample in cell cultures for isolation and amplification opf virus, which can take up to four days. The full four days are needed to confirm a negative result (two passages in cell culture each of 2 days duration) which means that descisions on disease control may have to be taken before laboratory test results are avaliable. In serious epidemics there is also the need to beable to process large numbers of samples.
More rapid and sensitive diagnostic procedures are therefore required to improve disease control and the reverse transcription polymerase chain reaction (RT-PCR) offers the potential to achieve this objective.
Universal and serotype specific primers will be investigated in a variety of formats to increase the reliability, scale-up potential and sensitivity of RT-PCR for the primary diagnosis of FMD and other vesicular viruses.
The project has the following objectives:
1. Evaluate TaqMan and SYBR Green RT-PCR methodology in combination with robotic techniques for FMD diagnosis using universal primers.
2. Develop "mimics" as controls to validate negative results in the TaqMan Rt-Pcr for FMD Diagnosis.
3. Evaluate TaqMan RT-PCR methodology for SVD diagnosis.
4. Validate procedures for the detection of FMD virus in milk and probang samples.
5. Design and evaluate alternative FMD primers for TaqMan RT-PCR methodology in combination with robotic techniques to facilitate consequent sequencing of the products.
6. Design and evaluate type-specific primers for serotype-specific tests for FMD diagnosis.
7. Analyse test line reactants on chromatographic (pen-side) strip tests by RT-PCR for confirmation of FMD diagnosis.
8. Evaluate Taqman Rt-PCR methodology for VSU diagnosis.
9. Evaluate TaqMan RT-PCR methodology for vesivirus diagnosis.
10. Evaluate "portable" PCR machines for FMD diagnosis.
Project Documents
• Final Report : Application of RT-PCR for the diagnosis of FMD and other vesicular diseases   (1001k)
Time-Scale and Cost
From: 2002

To: 2005

Cost: £540,893
Contractor / Funded Organisations
Institute for Animal Health (BBSRC)
Animal Diseases              
Animal Health              
Foot and Mouth              
Molecular Biology              
Plants and Animals              
Fields of Study
Animal Health