菠萝成分的实时荧光PCR检测方法的建立

Detection of pineapple ingredients by real-time fluorescence polymerase chain reaction

中文关键词:  食品  菠萝  实时荧光PCR

英文关键词:food  pineapple  real-time fluorescence polymerase chain reaction

中文摘要:

      目的 本文研究建立食品中菠萝成分的实时荧光PCR检测方法。方法 根据菠萝rbcL基因设计菠萝物种特异性检测引物和荧光探针, 对样品中的靶标基因片段进行检测, 并进行物种特异性检测、灵敏度测试和实际应用检测。结果 通过对供试的58种动植物材料进行检测, 只有菠萝出现特异性扩增, 其他物种材料无扩增; 对不同浓度菠萝DNA样品和不同含量的菠萝粉样品进行灵敏度测试, 该检测方法对菠萝成分的检测灵敏度分别为0.01 ng/μL菠萝DNA和0.1%菠萝粉; 对市场销售的实际样品进行检测, 能够满足于检测需求。结论 该方法简单、灵敏、快速、准确, 能应用于食品中菠萝成分检测。

英文摘要:

      Objective To establish a method for detection of pineapple ingredients in foods by real-time fluorescence polymerase chain reaction (PCR). Methods Specific primers and probe were designed according to highly conserved sequence of rbcL gene of pineapple. The target gene fragment in the sample was tested. The specificity, sensitivity and applicability of method for detection pineapple were carried out. Results The specificity of the method was verified by performing PCR experiments on 58 animals and plants. The methods were considered to be specific only from the pineapple and no amplified curves from other species. The limit of detection was 0.01 ng/μL pineapple DNA and 0.1% pineapple powder. The actual sample of the market were detected, it met the requirements of detection. Conclusion This method is simple, sensitive, fast and accurate, which can be used to the identification of pineapple ingredients in foods.