GMOs are organisms in which the genetic material has been altered in a way that does not occur naturally by mating and/or natural recombination. The European regulation in particular through the 2001/18/EC directive and both 1829/2003/EC and 1830/2003/EC regulations as regards traceability and labelling of GMOs (and derived products) in the agro-food sectors compels the Member States to have analytical tools for qualitative and quantitative detection of GMOs all along the food chain.
Objectives
The present project consists in extending the knowledge about GMO markers (and their collection) and improving the methods of screening, authentication and quantitation of GMOs in any foodstuffs. A first theme is the development of plasmids –as positive control for qualitative PCR or calibrators for quantitative PCR containing (event-specific) GMO markers and taxon markers. A second theme aims at elaborating a qualitative screening GMOChip. That is to say a biochemical matrix allowing to detect and identify a very large number of different GMOs mixed in a same product. This theme is an anticipation for the increase of diversity of marketed GMOs all over the world. The third theme consists in analysing the influence of the quality of a DNA preparation on quantitation of GMOs with the help of real time PCR. The aim is (to try) to define analytical parameters allowing to decide if a DNA extract is good or not for quantitation by real time PCR. Specific tasks are also devoted to the DNA stability and inhibitory effects of extracts on the PCR.
Results obtained
The main results obtained in this project are : Theme 1 : Setting up a bank of cloned GM and taxon markers, and an associated database. Comparison between plasmids (as calibrators for real time PCR) and reference materials issued from flours with certified (IRMM) GMO contents for the during the quantitation of GMOs by real time PCR. Theme 2 : Elaboration of a DNA chip using 3 consensus primer pairs for qualitative screening of 8 transgenic events and checking the specificity of the different used probes. Theme 3 : Characterization of DNA extracts stemming from different matrices by : - analysing the efficiency of amplification on soybean DNA. - bringing to light several types of inhibition of the PCR. - analysing the HPLC patterns from different kinds of extracts.
Contribution
The main tasks of the CRA-W in this project were the preparation of DNA master stock solutions for the realisation of the tasks allocated to the theme 3 (see objectives and results) and the analyses of the DNA extracts for the qualitative and quantitative characterization of these extracts. The DNA extracts produced and analysed here were performed from matrices which are commonly met in routine analysis and using different extraction methods (fractionated precipitation with organic solvents; use of silica resins and paramagnetic particles).
Partners
The project comprises five paid partners : - Institute of Public Health (ISSP) Service of Biosafety and Biotechnology Rue Juliette Wytsman 14, B-1050 Brussels. - Agricultural Research Centre (CLO) Department of Plant Genetics and Breeding Caritasstraat 21, B-9090 Melle - University Notre-Dame de la Paix (FUNDP) Unity of Cellular and Molecular Biology Rue de Bruxelles 61, B-5000 Namur - Veterinary and Agrochemical Research Centre (CERVA) Department Quality and Safety Leuvensesteenweg 17, B-3080 Tervuren - Walloon Agricultural Research Centre (CRA W) Department Valorisation of Agricultural products Chaussée de Namur 24, B-5030 Gembloux The network includes also no paid partners who are grouped together in a user committee. The coordination of the project is carried out by the Institute of Public Health (ISSP).
