芝麻物种特异性定性PCR方法的建立

Establishment of Specific Qualitative PCR Method for Sesame Species

为鉴别食品中芝麻成分的真实性,建立了一套芝麻物种特异性定性PCR方法。根据Sesamum indicum 2S albumin-like序列,设计了3对芝麻物种特异性定性PCR引物,并对引物的特异性、最适反应浓度和最适退火温度进行了筛选和优化,建立了芝麻物种特异性定性PCR检测方法。进一步对方法的灵敏度、检出限和对芝麻加工品的适用性进行了测试。结果显示,引物sesame-1F/1R的特异性最好,最优反应条件为引物浓度0.4μmol·L^(-1),退火温度60℃;该方法能够检测出芝麻质量分数为0.1%的样品,且对于芝麻油、芝麻酱2种性状的加工样品均有很好的扩增。该方法能够鉴别食品中芝麻成分,为食品质量安全监管提供了有力的技术支撑。

To identify the authenticity of sesame ingredient in food, a set of specific quantitative PCR method for sesame species was established in this study. According to the sequence of sesamum indieum 2S albumin-like sequence, three pairs of sesame species specific qualitative PCR primers were designed. Passed the primer specificity test, PCR reaction condition and system optimize, a sesame species-specific qualitative PCR detection method was established. Then we need to make further testing on sensitivity test, detection limit test and processed product test. The results showed that the specificity of the primer sesame-1F/1R was the best, and the primer concentration was 0.4μmol·L^-1and the annealing temperature was 60℃, which was the optimum reaction condition. The minimum detection limit of the specific PCR method was 0.1% （M/M）, two kinds of traits of processed samples such as sesame paste and sesame oil were well amplified. This method can identify the sesame ingredients in food which provides strong technical support for food quality and safety supervision.