关键词: 微流控技术, 现场快速检测, 实时荧光PCR, 转基因鉴定

Abstract:

Accurate identification and supervision of genetically modified （GM） crops are critical for biosafety assurance and seed industry development. Most existing detection methods rely on laboratory environments and fail to meet the practical requirements for rapid， on-site， and multi-target parallel identification. This study aimed to establish an integrated on-site rapid detection system that enables one-stop nucleic acid extraction， amplification and detection. Based on a handheld integrated GM on-site rapid detection device equipped with a high-monochromatic excitation light source， miniature spectrometer and semiconductor refrigeration module， supporting multi-target nucleic acid detection chips and sample processing tubes were developed via microfluidic technology and precision injection molding. The system achieves fully automated “sample-in， result-out” detection for plant seeds， leaves and other samples within 30 minutes， supporting simultaneous detection of up to seven targets with less than 1 minute of manual operation. The built-in battery allows eight independent detections on one full charge. In simulated samples， the limit of detection （LOD） for the MON810 event reached 0.1% （mass fraction） with no non-specific amplification. Reproducibility tests showed that the coefficient of variation （CV） of C_t values across different operators， instruments and test time points was less than 1.5%， and the results were highly consistent with those of conventional laboratory real-time PCR. This developed method enables accurate， rapid and portable on-site identification of GM ingredients， providing an efficient integrated solution for grassroots supervision and seed industry quality control.

Key words: microfluidic technology, on-site rapid detection, real-time PCR, GM identification