Development of a quadruplex-real-time-PCR for screening food for genetically modified organisms |
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Authors: | Thomas Gaudron Claude Peters Elodie Boland André Steinmetz Gilbert Moris |
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Affiliation: | (1) Laboratoire National de Santé 1A, rue Auguste Lumière, 1950 Luxembourg, Luxembourg;(2) Centre de Recherche Public de la Santé, 84 Val Fleuri, 1526 Luxembourg, Luxembourg |
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Abstract: | With the ever increasing number of genetically modified plants authorized worldwide, including in the European Union, high throughput detection methods need to be developed. In this paper, a quadruplex-real-time-PCR method is described which allows rapid and simultaneous screening of food for the presence of target DNA sequences from the cauliflower mosaic virus 35S promoter, the NOS terminator from Agrobacterium tumefaciens, the soya reference lectin gene and the maize reference alcool dehydrogenase gene (adh). Three of the four primers and probe combinations have already been published elsewhere, whereas primers and probe for NOS terminator-PCR were developed in-house. Validation data show sensitivities down to five copies for 35S promoter and NOS terminator PCR, even when target sequences of the competing PCRs are in large excess. Thorough adjustment of primer and probe concentrations allowed high individual PCR efficiencies with negligible physical cross-talk between the four detection channels. This method provides a basis for a rapid screening of food for the most frequently used regulatory elements present in GM crops authorized for food in the European Union. In addition it provides information about the presence of species which are possibly genetically modified. |
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Keywords: | Quadruplex real Time PCR Screening Genetically modified organisms 35S promoter NOS terminator |
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