Research on Rapid DNA Extraction Technology Compatible with Multiple Systems for Genetically Modified Maize (Zea mays)
XIAO Gong1,2, FU Li-Jin2,3, CHEN Xiao-Yun2, WANG Xiao-Fu2, PENG Cheng2, XU Jun-Feng1,2,*, YANG Lei2,*
1 College of Life Science, Zhejiang Normal University, Jinhua 321004, China; 2 State Key Laboratory for Quality and Safety of Agro-products/Key Laboratory of Traceability for Agricultural Genetically Modified Organisms, Ministry of Agriculture and Rural Affairs/Zhejiang Key Laboratory of Crop Germplasm Innovation and Utilization, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China; 3 College of Modern Agriculture, Zhejiang A&F University, Hangzhou 311300, China
Abstract：The commercialization of genetically modified (GM) maize (Zea mays) in China has accelerated significantly, creating an urgent need for rapid field detection technologies as conventional nucleic acid extraction and detection methods are often too complex and time-consuming for efficient screening. To develop an accurate and efficient on-site detection technology to enhance regulatory oversight, this study systematically evaluated 5 cell lysis methods for their compatibility with 3 detection platforms: Conventional PCR, quantitative real-time PCR (qPCR), and recombinase polymerase amplification (RPA), while also investigating the effects of high-temperature incubation on DNA extraction efficiency. Results demonstrated that the alkaline lysis method showed the best cross-platform compatibility, with rapid extraction reagent 1 performing optimally. Although high-temperature treatment significantly increased nucleic acid yield, it simultaneously caused the release of chlorophyll and other fluorescent substances that severely interfered with detection accuracy. Based on these findings, an integrated portable detection device combining quartz sand mechanical lysis, membrane filtration, and isothermal amplification technologies was developed. This system achieved complete sample-to-result detection within 25 minutes, representing a 6-fold efficiency improvement over conventional methods. Field validation confirmed 100% detection specificity for both DBN9936 GM maize event and Ruifeng125 GM maize event, with effective discrimination from non-GM crops and other transgenic varieties. In conclusion, this study successfully identified rapid extraction reagent 1 as a versatile extraction method compatible with multiple detection platforms, and innovatively developed a 'sample-to-result' on-site detection system that provides a rapid (<30 minutes), efficient solution for GM crop regulation.
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