Development and validation of multiplex GMO screening test in food, feed and seed

Genetically modified organism (GMO) is shortly defined as an organism that is obtained by gene transfer using modern biotechnologic approaches. The production platform and extent of GMOs have been increasing since 1996 when they were first produced. This increment has brought to attention many questions and concerns. As in the world, the GMOs issue occupy the agenda in Turkey. GMOs are highly discussed on media that attracts the attention and curiosity of the consumers. With the spread of GMOs, countries have started to create their own legislation. GMOs control analyzes have been carried out in private and public laboratories in Turkey since the enterance into force of the Biosafety Law No. 5977 in 2010. GMO screening analysis is the first step of the GMO control analysis. After that, if necessary, further analysis, i.e., quantity and typing analysis, is performed. Therefore, it is crucial to obtain accurate results from the screening analysis. Regions showing genetic modification in more than 90% of GMO products are 35S promoter, NOS terminator, FMV promoter regions. Commercial kits are also generally produced to scan these regions. Less further analysis is needed when these regions are scanned. The fact that these regions are scanned as multiplex in commercial kits provides practicality for the user. Therefore, it is preferred by laboratories. 35S promoter, NOS terminator, FMV promoter regions show the genetic modifications in more than 90% of GMO products. Commercial kits are mostly produced to scan these regions. Further analysis are rarely needed when these regions are scanned. Commercial kits provides practicality for the user by allowing multiplex scanning of these regions. Thus, these kits are preferred by the laboratories. On the other hand, commercial screening kits constitute the main cost item in GMO screening analysis. GMO screening kits used commonly in laboratories are imported which cause high cost of these analysis. The high analysis fees may cause customer complaints. This study aimed to create GMO screening test kit which had the same precision and accuracy with the commercial kit by obtaining required reactives separately and combining them under laboratory conditions. Multiplex test material was prepared as an in-house method to reduce the cost as much as possible. The test material sold as a commercial kit costs high fees as being ready to use. The necessity of preparation and usage of test chemicals in laboratory environment in order to reduce the high GMO analysis fees constituted the rationale of this study. As a result of this study, a multiplex GMO screening test with a LOD (limit of detection) level of 10 DNA copies was developed. Moreover, this multiplex GMO screening kit had a lower analysis cost compared to the commercial kit. It can be used in GMO test laboratories for routine GMO analysis studies since validation procedure is completed.