多重实时荧光PCR TaqMan-MGB杂交探针标记法快速鉴定转基因玉米MIR162

为对转基因玉米MIR162的进口实行监测、规范转基因玉米在国内市场中的流通，建立一种转基因玉米MIR162准确快速的检测方法，通过设计转基因玉米MIR162品系特异性序列的引物和探针，摸索和优化多重荧光PCR体系和参数，开发建立多重实时荧光PCR TaqMan-MGB杂交探针标记法快速鉴定转基因玉米MIR162。结果表明，转基因玉米MIR162品系标准品和平行样品中内标基因和品系特异性基因均出现明显扩增曲线，其他转基因玉米品系标准品仅内标基因有扩增曲线，空白对照无扩增曲线，说明该TaqMan-MGB探针对转基因玉米MIR162品系具有扩增特异性。该方法可作为鉴定筛查转基因玉米MIR162品系及其转化体成分的有效方法，用于规范管理转基因产品在市场上的流通。

Rapid Identification of Transgenic Maize MIR162 by Multiple Real-Time Fluorescent PCR with TaqMan-MGB Hybridization Probe Labeling

In order to monitor the import of transgenic maize MIR162 and standardize the circulation of transgenic maize in domestic market, an accurate and fast detection method for transgenic maize MIR162 was established. By designing primers and probes for the specific sequence of transgenic maize MIR162, the multiple fluorescent PCR system and parameters were explored and optimized, and a multiple real-time fluorescent PCR with TaqMan- MGB hybridization probe labeling method for rapidly identifying transgenic maize MIR162 was developed and established. The results showed that the internal standard gene and line-specific gene of transgenic maize MIR162 line standard and parallel samples presented obvious amplification curves, and only the internal standard gene of other transgenic maize line standards exhibited amplification curves, while the blank control had no amplification curves, indicating that the TaqMan-MGB probe had amplification specificity for transgenic maize MIR162 strain. This method can be used as an effective method to identify and screen transgenic maize MIR162 strains and transformant components, and can be applied for regulating the circulation of transgenic products in the market.