New trait-specific qualitative SYBR®Green qPCR methods to expand the panel of GMO screening methods used in the CoSYPS

摘要

Since 2011, a new Commission Regulation (EU/619/2011) defines that laboratories testing for genetically modified organisms (GMO) need to be able to detect also genetically modified (GM) events pending for authorisation. This, in addition to the fact that the number of GM events authorised in the European Union (EU) that need to be identified multiplies rapidly and that the detection of unauthorised GMO becomes more important, led to the development of a time and cost-effective screening approach. Moreover, the GM elements that are utilised in the transgenic inserts also become increasingly diverse. Consequently, the screening approaches have to be updated to enable full coverage and better discrimination of all these events. To respond to this need, two new qualitative SYBR®Green real-time PCR (qPCR) methods were developed and in-house validated: one method is element-specific and targets the Cry3Bb trait, and the other one is a construct-specific method detecting the gat-tpinII junction. Method acceptance parameters such as the sensitivity, specificity and repeatability were assessed as well as the robustness of the methods. Additionally, the reproducibility was evaluated by transferring the methods to a second laboratory. Both methods allow a specific, sensitive and repeatable detection of the respective targets in food and feed samples and can be easily applied in a routine laboratory. Moreover, they can be used together with previously validated SYBR®Green qPCR methods to expand the panel of screening methods. This allows an extended coverage of the GM events authorised in the EU and adds discriminative power to the screening phase.