Theme 4 “Diagnostics”

Theme leader: Dr Martin Beer, Friedrich-Loeffler-Institute, Germany.

Deputy Theme leader: Dr Åse Uttenthal, National Veterinary Institute, Technical University of Denmark, Denmark.

Prevention and control of epizootic diseases starts with the use of diagnostic tools to detect infected animals. EPIZONE has the intention to standardise and optimize (new) diagnostic methods and their use in prevention and control of epizootic diseases.

Work Package 4.1: PCR-diagnostics
Work Package 4.2: DNA-Chip-based diagnostics
Work Package 4.3: DIVA Diagnostics
Work Package 4.4: Pen-side tests

What is going on in the EPIZONE network in the field of Diagnostics

·          Harmonization and standardization activities of real-time RT-PCR assays are performed for foot-and-mouth disease virus (FMDV), classical swine fever virus (CSFV), avian influenza virus (AIV), Newcastle disease virus (NDV) and bluetongue virus (BTV) (WP 4.1).

·          A ring trial succeeded in the optimization of CSFV-specific real-time RT-PCR diagnostics in the EU. Laboratories with a sub-optimal protocol changed to the highly sensitive and specific standard method or improved their own protocol (WP 4.1).

·          A first CSFV-RNA reference panel is distributed. This so-called “EPIZONE REFERENCE RNA PANEL CSFV” can be used by all EPIZONE partners for a further harmonization of CSFV real-time PCR diagnostics, DNA-Chip diagnostics  as well as pen-side testing (WP 4.1).

·          A unique pan viral DNA chip and DNA chips for accurate sub-typing of important epizootic viruses are designed. A combined “EPIZONE Pan Viral DNA Chip” is under evaluation and will be tested during workshops and in a ring trial (WP 4.2).

·          Information on DIVA diagnostics in CSFV, FMDV and HPAI is collected in three different reviews. A paper based on those reviews is in preparation (WP 4.3).

·          Three international workshops have been conducted involving national crisis management personnel from EPIZONE and non-EPIZONE partners. The workshops have discussed the use and interpretation of DIVA diagnostics for foot-and-mouth disease after vaccination in domestic herds (WP 4.3).

·          The interaction of experts of EPIZONE on “pen-side tests” is enhanced. As a main result, the loop mediated amplification methods (LAMP) appear to be emerging as a front-runner and focal point for further investigation by the EPIZONE partners. In addition it was considered useful to include the participation of commercial partners in the area of point of care testing in discussions (WP 4.4).

Publications

WP 4.1

Catherine Cetre-Sossah, Bruno Mathieu, Marie-Laure Setier-Rio,  Colette Grillet, Thierry Baldet, Jean-Claude Dele´colle, Emmanuel Albina; “Development and evaluation of a real-time quantitative PCR assay for Culicoides imicola, one of the main vectors of bluetongue (BT) and African horse sickness (AHS) in Africa and Europe”. Research in Veterinary Science 85 (2008) 372–382.

WP 4.2

Peter Gyarmati, Tim Conze, Siamak Zohari, Neil LeBlanc, Mats Nilsson, Ulf Landegren, Johan Baner and Sandor Belak (2008). “Simultaneous Genotyping of All Hemagglutinin and Neuraminidase Subtypes of Avian Influenza Viruses by Use of Padlock Probes”. Journal of Clinical Microbiology, 2008, 1747–1751.

Astrid Gall, Bernd Hoffmann, Timm Harder, Christian Grund and Martin Beer(2008). “Universal primer set for amplification and sequencing of HA0 cleavage sites of all influenza A viruses”. Journal of Clinical Microbiology, 46, 2561-2567.

WP 4.4

Michaud et al., (2007) “Long-term storage at tropical temperature of dried-blood filter papers for detection and genotyping of RNA and DNA viruses by direct PCR”. Journal of Virological Methods, 146, 257-265.