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A Microarray-based Detection System for Genetically Modified (GM) Food Ingredients
- Leimanis, S., Hernandez, M., Fernandez, S., Boyer, F., Burns, M., Bruderer, S., Glouden, T., Harris, N., Kaeppeli, O., Philipp, P.
- Plant molecular biology 2006 v.61 no.1-2 pp. 123-139
- food crops, recombinant DNA, microarray technology, transgenic plants, Agrobacterium radiobacter, polymerase chain reaction, kanamycin kinase, Streptomyces hygroscopicus, Cauliflower mosaic virus, reporter genes, promoter regions
- A multiplex DNA microarray chip was developed for simultaneous identification of nine genetically modified organisms (GMOs), five plant species and three GMO screening elements, i.e. the 35S promoter, the nos terminator and the nptII gene. The chips also include several controls, such as that for the possible presence of CaMV. The on-chip detection was performed directly with PCR amplified products. Particular emphasis was placed on the reduction of the number of PCR reactions required and on the number of primers present per amplification tube. The targets were biotin labelled and the arrays were detected using a colorimetric methodology. Specificity was provided by specific capture probes designed for each GMO and for the common screening elements. The sensitivity of the assay was tested by experiments carried out in five different laboratories. The limit of detection was lower than 0.3% GMO for all tests and in general around 0.1% for most GMOs. The chip detection system complies with the requirements of current EU regulations and other countries where thresholds are established for the labelling of GMO.