An innovative integrated approach based on DNA walking to identify unauthorized GMOs

Marie-Alice Fraiture, Philippe Herman, Isabel Taverniers, Marc De Loose, Dieter Deforce, Nancy H Roosens

    Onderzoeksoutput: Bijdrage aan congresGepubliceerd abstract

    Uittreksel

    In the next coming years, the frequency of unauthorized genetically modified organisms (GMOs) being present in the European food/feed chain will increase significantly. Rice already constitutes a challenge for laboratories developing methods to detect unauthorized GMOs. Indeed, in 2012, several genetic modified rices were detected in products imported from Asia, mainly from China. Therefore, we have developed a strategy to identify unauthorized GMOs containing a pCAMBIA family vector, frequently present in transgenic plants. The presented integrated approach is performed in two main successive steps on Bt rice grains. First, the potential presence of unauthorized GMOs is assessed by the qPCR SYBR®Green technology targeting the terminator 35S (t35S) pCAMBIA element, which allows discriminating pCAMBIA family vectors. Second, its presence is confirmed via the characterization of the junction between the transgenic cassette and the rice genome. To this end, a DNA walking strategy is applied using a first reverse primer followed by two semi-nested PCR rounds using primers that are each time nested to the previous reverse primer. The sensitivity of the method was assessed. This innovative approach allows to rapidly identifying the transgene flanking region and presents the advantage to be easily implementable in GMO routine analysis by the enforcement laboratories.
    TaalEngels
    StatusGepubliceerd - 2013
    EvenementTrends in Food Analysis VII - Gent, Gent, België
    Duur: 19-sep-201319-sep-2013

    Symposium

    SymposiumTrends in Food Analysis VII
    LandBelgië
    StadGent
    Periode19/09/1319/09/13

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