TaqMan实时荧光PCR检测菰米成分的方法建立

本研究建立了一种能够快速检测食品中菰米成分的TaqMan实时荧光PCR方法。以菰米核糖体内转录间隔区（internal transcribed space, ITS）基因为检测靶标，设计特异性引物和探针，建立菰米成分的实时荧光PCR检测方法，并进行物种特异性分析、灵敏度分析以及实际应用检测。结果显示:本研究所建立的实时荧光PCR检测方法特异性强，仅对菰米品种中国菰（Z. latifolia）、水生菰（Z. aquatica）、沼生菰（Z. palustris）和德克萨斯菰（Z. texana）基因组DNA出现特异性扩增曲线，供试的其他30种谷物以及异源性动植物基因组DNA均无扩增曲线；该方法对菰米成分检测的灵敏度为0.001 ng/μL菰米基因组DNA或0.01%（W/W）菰米粉。应用该方法对100份市售的进口菰米、菰米碎米、杂粮米及混合米粉等样品进行检测，在80份进口菰米和5份菰米碎米中检测出菰米成分，其他样品中未检出菰米成分，与商品标识一致。该方法灵敏度高，特异性强，可快速高效地进行菰米成分的真实性甄别。

Establishment of TaqMan Real-time PCR Method for Detection of Wild Rice Derived Ingredients

In this study, a TaqMan real-time PCR method for rapid detection of wild rice in foods was established. Specific primers and probes were designed according to conserved sequence of internal transcribed space (ITS) gene of Z. latifolia, Z. aquatica, Z. palustris and Z. texana to test the target gene fragment in the samples, and then species-specific detection, sensitivity detection and practical application detection were carried out. Results showed that this real-time PCR method had strong specificity, only showed specific amplification curves for wild rice genomic DNA, and there was no amplification curve for other grains, animal and plant materials. The limit of detection was 0.001 ng/μL genomic DNA or 0.01% (W/W) wild rice powder per reaction. The feasibility of the method was further verified by detecting 100 samples of commercial imported wild rice, broken wild rice, coarse grain and rice flour. The wild rice ingredients were detected in 80 commercial imported wild rice and 5 broken wild rice samples. This method has the characteristics of strong specificity, high sensitivity, rapidity and high sufficiency, and is suitable for rapid identification of wild rice ingredients.