Title: Development and validation of new diagnostic tools to equip the UK National Reference Laboratory for footandmouth disease.
Funder: Department for Environment, Food and Rural Affairs

Abstract: Description Foot-and-mouth disease (FMD) is difficult and expensive to control since the causative virus (FMD virus: FMDV) is highly contagious and spreads rapidly through susceptible livestock (cattle, sheep, goats and pigs). FMD is endemic in many countries in Africa and Asia and is globally one of the most widespread epizootic animal diseases. Since 2010, there has been an upsurge in cases of the disease in regions neighboring Europe (such as North Africa and The Middle East), as well as in countries with strong trade links to the UK (such as China). The elevated circulation of FMDV in these endemic regions inevitably poses increased threats for onward spread to FMD-free countries. In the event of an incursion, timely and accurate laboratory diagnosis is a vital component of prompt disease reporting and the control of FMD outbreaks. Tests are used to (i) confirm that FMDV is present on farms, (ii) to characterize circulating field strains of the virus and to differentiate from other viruses causing vesicular diseases, and (iii) to recognize serologically positive animals that have been exposed to FMDV. The focus of this new three-year project is to develop and validate enhanced virological and serological tests for the diagnosis of FMD, to ensure that the FMD National Reference Laboratory (NRL) at Pirbright (and surge capacity at APHA Weybridge) is equipped to respond rapidly to any future threats of FMD in the UK. The objectives of this project address the priorities of the NRL for FMD, and will deliver (i) new molecular and serological tests that can recognize FMDV infected animals at the earliest stages of disease, (ii) enhanced capability to respond to new threats (such as novel antigenic variants of FMDV and an emerging virus [Seneca Valley virus] that causes vesicular disease in pigs), (iii) resilience in the testing systems via the adoption of alternative serological test kits, as well as new assays that are not dependent upon high-containment facilities, (iv) serological tests that can be used to identify animals that have been previously exposed to FMD, particularly those within a vaccinated population according to the EU Directive and Recommendations of OIE, and (v) validated cost-effective kits and reagents for high through-put FMD diagnosis. These tests can be used for rapid identification of infected animals as the basis for more targeted control, with a consequent reduction in the costs of FMD outbreaks (industry and government), and to provide an evidence-base to support a future reduction in the length of time businesses remain under official restrictions. This work will also contribute to maintaining the Quality Assurance framework for these diagnostic tests, and provides trained scientists and diagnostic expertise that can be employed in the event of a future FMD outbreak in the UK. Objective Work undertaken within this project will fall into the following objectives:1: Validation and accreditation of alternative cell line(s) that can be used to culture FMD virus2: Adopt a newly developed antigen-detection ELISA (from IZSLER/Pirbright) for routine diagnostic use3: Develop and validate additional reagents for use in new Ag-ELISA formats (including those that can be generated without the need for high-containment facilities)4: Modification of real-time RT-PCR protocol to use less expensive master-mix reagents5: Validate and adopt a real-time RT-PCR assay for the detection of SVV-16: Compare the analytical sensitivity between different amplification technologies that might be selected for rapid “field” detection assays7: Adopt alternative NSP ELISA kit into the portfolio of ISO/IEC 17025 accredited tests8: Validate multiplex NSP Luminex test for the detection of FMDV-specific antibodies9: Develop and validate additional reagents for use in new in-house SP ELISA formats (including those that can be generated without the need for high-containment facilities)10: Prepare validation dossiers for commercially available SP ELISAs for serotypes A and Asia-111: Validate IgA ELISAs that employ recombinant capsids for the detection of sub-clinical infection in cattle and small ruminants (serotypes O, A and Asia-1)12: Validate IgM ELISAs for early detection of FMDV-specific antibodies in cattle and sheep


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: INFECTION