Real-time PCR methods for the detection of DNA constructs with the nptII gene for the detection of genetically modified plants in food, feed and seed

摘要

For routine analysis of the most different food and feed matrices for genetical modification screening methods have increasingly been applied during the past years as the first step of detection. Screening for frequently used regulation elements and recombinant DNA-constructs by the use of real-time PCR is the state of the art. By combining several screening methods, the number of lines of genetically modified plants existing in one sample can be delimited. Apart from the genetically modified plants authorized in the EC, for which reference material and detection methods have to be made available by the placer on the market, there has been an increase of unauthorized GMO on the European market recently, for which event specific detection methods are not available as a rule. It does therefore have to be the target of routine controls by the laboratories charged with surveillance to be able to detect as much as possible of genetically modified plants with the aid of just a few screening PCR methods. Three different construct specific real-time PCR methods are presented in this study. They amplify the junction sequence from the 35S-promoter or the nos-promoter to the nptII gene. Furthermore, a gene specific real-time PCR method for the detection of DNA sequences of the nos-promoter from Agrobacterium tumefaciens is presented.