Application of Digital PCR and Real-time PCR on Screening Detection of GM Components of Soybean Milk
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Abstract:
In order to solve the implementation problems of current GM food standards and improve the inspection and supervision of GM foods in China, the application of digital PCR (dPCR) in the screening detection of genetically modified organisms (GMOs) was developed. In this study, the transgenic promoter CaMV35s and the terminator NOS genes were selected as targets and soybean endogenous gene Lectin as internal standard, and the detection limit were determined using transgenic soybean standard on real-time PCR (RT-PCR) and ddPCR systems. Two methods were both applied to GM screening detection for 30 batched of soybean milk. The results showed that the concentration detection limit of ddPCR reached 0.04 ng/reaction, and the limit of content detection reached 0.05%, which were both superior to RT-PCR (0.2 ng/reaction, 1%). In addition, 0.05% was lower than the most stringent labeling threshold level 0.9% of the European Union (EU). In the practical application of soybean milk, 26 batches of samples were consistently negative and 1 was positive on both of the two platforms, while 3 batches of samples with Ct values between 34.59~38.28 were confirmed by ddPCR, indicating that dPCR could help confirm the uncertain results of RT-PCR and slove the current standard judgment problems.
