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Content DescriptionThis document specifies a procedure for the detection of a DNA sequence of the open reading frame five (ORF V) from cauliflower mosaic virus (CaMV) and a procedure for the detection of the DNA sequence of the nopaline synthase (nos) gene from tumour-inducing (Ti) plasmids of phytopathogenic Rhizobium radiobacter (formerly named Agrobacterium tumefaciens). The procedures can be used in the context of screening for genetically modified crop/plants and their derived products to further clarify a positive PCR result for a specific promoter or terminator of CaMV (P-35S, T-35S), or both, and the nos gene (P-nos, T-nos), respectively. The methods specified in this document will detect and identify naturally occurring CaMV or Rhizobium radiobacter (Ti plasmid) DNA, or both, if present in the sample in the absence of a genetically modified plant event containing the specified target sequences. Both methods are based on the real-time polymerase chain reaction (PCR) and are applicable for the analysis of DNA extracted from foodstuffs and other products such as feedstuffs and seeds/grains. The application of the methods requires the extraction of an adequate amount of amplifiable DNA from the relevant matrix. With appropriate calibration material, the CaMV ORF V or nos copy number, or both, can be estimated and compared, respectively, with the estimated copy number for the promoter (P-35S, P-nos) or the terminator (T-35S, T-nos) sequences, or both. Thereby, conclusions are possible about the presence of an unknown genetically modified organism (GMO) in addition to any detected CaMV DNA or Rhizobium radiobacter Ti plasmid DNA, or both, in a test sample. About ISOISO, the International Organization for Standardization, brings global experts together to agree on the best way of doing things – for anything from making a product to managing a process. As one of the oldest non-governmental international organizations, ISO has enabled trade and cooperation between people and companies all over the world since 1946. The International Standards published by ISO serve to make lives easier, safer and better. |
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