转基因检测微流控恒温扩增芯片的研制

近年来环介导等温扩增技术（Loop-Mediated Isothermal Amplification，LAMP）越来越多地应用于转基因农作物、微生物等的基因检测中。为了提高相关检测机构的转基因产品检测效率，加快进出口食品农产品的通关速度，本文首次使用碟式芯片环介导恒温扩增技术，在65 ℃恒温条件下，对样品中的P-CaMV35S、T-NOS、NPTⅡ、BAR、PAT、Cry1Ac、EPSPS I、EPSPS II、FMV35S和GM-HRA等转基因靶标进行同步检测，检出限可达到0.5%（m/m），实现一次上样即可完成样品的转基因成分筛查。该方法特异性和灵敏度良好、检测时间较短，可实时读取扩增结果。对实际样品的检测结果表明，该方法的检测结果与实时荧光PCR方法的完全一致。本文开发的转基因检测微流控恒温扩增碟式芯片，可用于常见农产品（大豆、玉米、大米和棉花等）的转基因成分筛查，为转基因产品的定性检测提供了新的快速、灵敏、高通量技术平台。

Development of a Microfluidic Chip for Detection of Genetically Modified Organisms by Loop-mediated Isothermal Amplification

Loop-mediated isothermal amplification (LAMP) has been increasingly applied in the genetic detection of transgenic crops and microorganisms in recent years. In order to improve the detection efficiency of related inspection agencies for transgenic products and speed up the clearance of imported and exported agricultural products and foods, LAMP was combined with a microfluidic chip for the first time to simultaneously detect ten genetic recombinant elements, including P-CAMV35s, T-NOS, NPT, BAR, PAT, Cry1Ac, EPSPSI, EPSPSII, FMV35s, and GA-Hra, in samples at 65°C. The limit of detection could reach 0.5% (m/m), and the detection of transgenic ingredients in the sample could be completed with one sample loading. This method had a good specificity, a high sensitivity, and a short detection time, and the amplification signals could be acquired in real time using software. The detection results for actual samples indicated that all the results were consistent with those obtained by the real-time polymerase chain reaction (PCR) standard method. The microfluidic chip developed in this study for LAMP can be used for the inspection of transgenic ingredients in common agricultural products (soybean, corn, rice, cotton, etc.), and provides a rapid, sensitive, and high-throughput technical platform for the qualitative detection of transgenic products.