Ultrasensitive and visual detection of genetically modified crops using two primers-induced cascade exponential amplification assay

The increased global cultivation area of genetically modified (GM) crops has caused severe controversies over potential health and environmental risks worldwide. There is an urgent need to verify even trace amount of a particular GM material in products. Herein, a two primers-induced cascade exponential amplification reaction combined with cationic conjugated polymers (CCPs)-based visual detection method is developed for rapid and ultrasensitive detection of GM crops. This method only uses two primers to specifically recognize the four regions of the target gene, which is easier for primer design and probably more suitable for the detection of shorter targets. By integrating the two exponential amplification reactions, as low as 5 pg genomic DNA from GM maize can be accurately detected, which is more sensitive than the single amplification-based methods. Taking advantage of the efficient fluorescence resonance energy transfer (FRET) between CCPs and the commercial fluorescent dye SYBR Green I (SG), our method can differentiate as low as 0.01 % GM maize from a large amount of non-GM maize, which is the most accurate method so far. By changing the two primers according to target gene, our method can be modified to the detection of any other GM materials, indicating that our method is promising to be applied in other GM materials-related testing and screening system.