Abstract:

Test kits based on novel antibody fusion proteins (AFP) will be produced for the rapid, reliable and harmonised detection of three plant viruses of statutory or quarantine importance within the European Union. The AFP will be obtained by recombinant DNA methods, and large quantities will be produced in vitro using heterologous expression systems. They will be incorporated into prototype self-contained test kits. The composition of the kits will be optimised for stability and robustness. AFP test kits will be provided to national testing services and agencies for validation. The results will be widely disseminated at meetings and in journals of appropriate scientific societies, plant health agencies, growers and trade organisations.

Objectives:

The aims of the project are to produce and validate test kits based on antibody fusion proteins (AFP) to detect and identify three harmful viruses - tomato spotted wilt virus (TSWV), potato leafroll virus (PLRV) and beet necrotic yellow vein virus (BNYVV) - and to prove the benefits on a realistic scale. The target viruses are of statutory or quarantine significance and pose a threat to the free movement of plants and plant propagation material in Europe. Currently, laboratories obtain reagents for these viruses from a variety of different sources. With the introduction of plant passports it is important to have confidence in the results of tests done across the European Union. Therefore, standardisation of tests and harmonisation of test methods is desirable.

Yields of functional AFP produced in Escherichia coli, Drosophila and Pichia expression systems will be compared in pilot expression studies. Large quantities of AFP will be produced in the best systems and purified. Prototype kits will be designed and components optimised for stability and performance. The kits will be extensively evaluated by end-users and modified as necessary.

Once the effectiveness of both the new technology and the reagents have been demonstrated, the relevant plant health authorities and technology users will be notified and we expect the results to influence policy and assist harmonisation of tests for these viruses.

Contacts

Coordinator

• Lesley TORRANCE / Scottish Crop Research Institute UNITED KINGDOM

EC Scientific Officer

• European Commission DG Agriculture BELGIUM

Participant

• Bernard MOLLARET / BIORAD FRANCE
• Uli COMMANDEUR / RWTH, Aachen University of Technology Institute for Biology VII, Molecular Biotechnology / GERMANY
• Florian RUKER / University of Ag Insitute of Applied Microbiology / AUSTRIA
• Arjen SCHOTS / Wageningen Agricultural University Lab. of Nematology / NETHERLANDS