WP8 Confirmation and reference techniques


The overall objective of this WP is to develop and implement techniques for confirmatory reasons and as a basis
for adaptations especially of molecular tools.

The detailed aims of this Work package are the following:

  1. To develop high technology strategies that can be applied to confirm the status of a sample irrespective of gold standard technologies, without prior knowledge. At the genome detection level: identification of sample status without prior knowledge. Methods will include sequencing (with sequence independent access methods, including next generation sequencing with 454/Roche technology) and highly multiplexed genome detection methods including microarray systems. These technologies will be of use in diagnostically difficult situations like (re-)emergence of unexpected pathogens or pathogen variants (including “diagnostic escape mutants”).
  2. To improve our sequence knowledge with aim to improve test design and evaluation. To achieve this: development of more efficient and economic methodologies to sequence viral genomes (“towards the 100 euro viral genome?”) and their application to generate sequence information. Economic and efficient access to sequence information will enhance on the long term the available knowledge in public databases needed for efficient design and evaluation of molecular diagnostics.
  3. To use multiplex immunoassay for viral diagnosis with a Luminex platform for multiplexed Ab detection. The design of the method here would not be sequence/pathogen independent. The available reagents (antigens, monoclonal antibodies) will be collected from the partners and used to develop the confirmatory technology.

Multiplex immunoassay for viral diagnosis (e.g. FMDV, BTV, West Nile Virus) by using the Luminex liquid array technology will be developed/tested and used as confirmatory technology. This assay will allow e.g. the simultaneous detection of antibodies or antigens, in a single reaction, to multiple viral antigens coupled to microspheres labelled with different proportions of fluorescent dyes.

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