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Application of RT- PCR for the diagnosis of FMD and other vesicular diseases - SE1121

Description
Rapid and accurate laboratory detection of foot-and-mouth disease virus (FMDV) plays a critical role in the implementation of control measures to eradicate FMD. We have developed an automated RT-PCR assay that can be used to complement existing diagnostic methods such as virus isolation and antigen ELISA to detect FMDV. Assays in a similar format have also been developed to detect other vesicular disease viruses such as those of swine vesicular disease which can be confused with FMD. We have shown that these assays are highly sensitive and specific, and can be successfully applied to the full variety of sample types that can be received at the laboratory. The high potential through-put of samples achieved by automated protocols within a typical working-day compares favourably with virus-isolation methods, which is also highly sensitive, but can take up to 4 days to produce a definitive result.In view of the potential benefits of RT-PCR as a diagnostic tool, there has been increasing acceptance of these methods for routine use. This project aims to build on our previous work in this field, to further define and improve the performance of the RT-PCR, and to explore novel and alternative approaches that may become useful for FMDV and other vesicular disease virus detection. This 3-year project consists of the following objectives:· An inter-laboratory proficiency test involving national reference laboratories will be co-ordinated by IAH. This exercise will compare the performance of the Pirbright automated real-time RT-PCR assay with other molecular assays. Material suitable for use as standards for RT-PCR methods will be prepared and tested.· An encapsulated internal control to validate negative results will be developed for use in the TaqMan® RT-PCRs for FMDV and other vesicular disease viruses.· Multiplex assays will be developed that are capable of the simultaneous detection of a variety of PCR targets. These assays will be used for differential diagnosis of FMD from other vesicular disease viruses, and to provide serotype-specific and other characterisation data for FMDV isolates.· We will develop a modified single-tube RT-PCR that will be compatible with mobile PCR equipment that could be used in the field or located in regional laboratories. Since this technology is designed for use by non-laboratory personnel, these assays have the potential to rapidly deliver highly sensitive diagnostic results rapidly close to the suspect case of FMDV.· The suitability of alternative amplification and immunocapture template enrichment strategies will be investigated. These formats may be particularly suited for use in mobile/field-based situations (or in the developing world), where access to “fragile” precision equipment is not practical (or too expensive). Together, these objectives will aim to fully realise the potential of molecular methods such as RT-PCR for use in the detection of FMDV and other vesicular disease viruses.
Project Documents
• Final Report : Application of RT- PCR for the diagnosis of FMD and other vesicular diseases   (2833k)
Time-Scale and Cost
From: 2005

To: 2008

Cost: £585,405
Contractor / Funded Organisations
Institute for Animal Health (BBSRC)
Keywords
Animal Diseases              
Animal Health              
Biotechnology              
Diagnosis              
Foot and Mouth              
Plants and Animals              
Fields of Study
Animal Health