TY - Generic T1 - An innovative integrated approach based on DNA walking to identify unauthorized GMOs Y1 - 2013 A1 - Marie-Alice Fraiture A1 - Philippe Herman A1 - Taverniers, Isabel A1 - De Loose, Marc A1 - Nancy Roosens KW - An innovative integrated approach based on DNA walking to identify unauthorized GMOs AB -

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.

PB - Trends in Food Analysis VII Symposium CY - Gent, Belgium ER -