植物蛋白饮料含有丰富的营养成分,深受广大消费者的喜爱。但由于利益驱使,掺假现象也逐年增加。针对此现象构建了核桃、花生、大豆3种植物成分的阳性质粒分子,对3种阳性质粒分子的特异性和灵敏度进行了验证,并对实时荧光PCR反应体系中不同品牌PCR预混液效果进行了对比分析。结果表明,构建的核桃、花生、大豆阳性质粒分子均具有特异性强、灵敏度高等特点,检验灵敏度均可达到10-5 ng/μL;标准曲线扩增效率分别为93.299%、123.623%和132.975%,且相关线性系数R2均在0.99以上。综上所述,3种品牌的PCR预混液均具有很高的灵敏度,均可以满足核桃、花生和大豆日常检验工作的要求,对于植物源性成分的鉴定具有重要意义。
Vegetable protein beverages are popular among consumers because of their rich nutrients. However, adulteration has increased year by year due to the pursuit of profits. In response to this phenomenon, three positive plasmid molecules for detection were constructed in this paper. Three plant constituents of positive plasmids (walnut, peanut and soybean) were constructed, the specificity and sensitivity of three positive plasmids were verified, and the effects of different PCR premixed liquid on the real-time fluorescent quantitative PCR reaction were compared and analyzed. Results showed that the constructed positive plasmid molecules of walnut, peanut and soybean had strong specificity and high sensitivity, and the test sensitivity could reach 10-5 ng/μL. The amplification efficiency of the standard curve is 93.299%, 123.623% and 132.975%, respectively, and the correlation coefficients R2 are all above 0.99. In summary, the three commercial PCR premixed liquid have high sensitivity, which could meet the requirements of routine detection of walnuts, peanuts and soybeans. It is of great significance for the identification of plant-derived components.
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