Volume 2 Article informations A new Multiplex PCR-based strategy to detect GM maize events with 2k fractional factorial plan in processed food Nesrine Nabi1, Mohamed Salem Zellama1, Ahmed Ben Hafsa1 and Maher Chaouachi 1,* Abstract Governments in many countries have established a policy of labeling all food and feed containing or produced by Genetically Modified Organisms (GMO). Consequently, versatile, laboratory transferable GMO detection methods are in increasing demand. Here we describe a new 2K fractional factorial plan of optimization approach for multiplex polymerase chain reaction (PCR) system used for the simultaneous detection of target sequences in genetically modified maize (MON810, Bt176, and T25). Primer pairs were designed to amplify the junction regions of the transgenic constructs analyzed and the endogenous genes of maize (Adh1) were included as internal control targets to assess the efficiency of all reactions. These results demonstrated the specificity, reproducibility and flexibility of this process and indicated that this multiplex PCR method using the 2K Fractional Factorial Plan could be an effective qualitative detection assay for screening GM maize. Here, we describe an experimental design to attain easier, quicker and cheaper PCR-based assay optimization, suitable for the detection of GM maize events in a single reaction. Key words: GMO detection, Fractional factorial plan, optimization, PCR, regulation, validation.