Abstract
The presence of genetically modified organisms (GMOs) in food and feed products is regulated in many countries. Various countries have issued labeling threshold levels to supervise GMO commercialization, and efficient detection methods to quantify GMO contents are critical to meet labeling regulations. The aim of this study was to develop event-specific methods to measure the event-specific/lectin gene (Le1) copy number ratio in soybean using droplet digital PCR (ddPCR), which is a recently developed technology that offers high accuracy. Duplex ddPCR methods were developed and optimized using a central composite design for the five most widely authorized GM soybean events, in which the target gene was labeled with one fluorescent reporter and the endogenous gene with another. The methods produced specific results, and the performance complied with international GMO labeling regulations. Further, the duplex ddPCR methods are compatible with chip-based digital PCR platforms and original real-time PCR, but allow higher throughput and are more sensitive in identification and quantification of these five widely authorized genetically modified soybean events.
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Acknowledgements
The Chinese Ministry of Agriculture (Changchun) is acknowledged for providing and validating the blinded samples of the target GM soybean events.
Funding
This research was supported by the Jilin Agricultural Technological Innovation Project, China (CXGC2017JQ017), and the National Transgenic Plant Special Fund, China (2018ZX08012001).
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F.L., as project leader, designed this study. L.L. and W.Y. performed the main laboratory work and wrote the manuscript. Y.H., L.D., and Z.X. performed some laboratory work. C.L. and W.X., as supervisor, provided direction. All authors contributed to discussions, and read and approved the final manuscript.
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Feiwu Li declares that he has no conflict of interest. Likun Long declares that she has no conflict of interest. Wei Yan declares that she has no conflict of interest. Yuxuan He declares that she has no conflict of interest. Liming Dong declares that he has no conflict of interest. Zhenjuan Xing declares that she has no conflict of interest. Congcong Li declares that he has no conflict of interest. Wei Xia declares that she has no conflict of interest.
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Likun Long and Wei Yan contributed equally to this work and are co-first authors.
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Long, L., Yan, W., He, Y. et al. Development of a Duplex Digital PCR Method to Quantify Five Genetically Modified Soybean Events. Food Anal. Methods 15, 294–306 (2022). https://doi.org/10.1007/s12161-021-02104-2
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DOI: https://doi.org/10.1007/s12161-021-02104-2