五重实时荧光聚合酶链式法快速筛查食用淀粉及其制品中植物源性成分

Rapid screening of plant-based components in edible starch and its products by five fold real-time fluorescence polymerase chain reaction

中文关键词:  多重实时荧光PCR  食用淀粉  淀粉制品  豌豆  绿豆  木薯  玉米

英文关键词:five fold real-time fluorescence polymerase chain reaction  starch  starch products  detection method

中文摘要:

      目的 建立五重实时荧光聚合酶链式方法快速筛查食用淀粉及其制品中植物源性成分。方法 首先设计豌豆、绿豆、木薯、玉米的特异性引物、探针, 建立多重实时荧光聚合酶链反应(polymerase chain reaction, PCR)检测方法, 再验证方法的特异性和绝对灵敏度, 最后通过样品掺假模拟实验验证方法的检出限。结果 该方法的特异性强, 仅能检出豌豆源性、绿豆源性、木薯源性、玉米源性成分; 灵敏度较高, 对绿豆源性和木薯源性的检测灵敏度可达到1×10–1 ng/μL水平, 豌豆源性和玉米源性的检测灵敏度可达到1×10–2 ng/μL水平。结论 本研究建立的五重实时荧光PCR方法可用来区分市场上淀粉及其制品中的豌豆源性、绿豆源性、玉米源性和木薯源性成分, 具有较好的应用价值。

英文摘要:

      Objective To establish a five fold real-time fluorescence polymerase chain reaction (PCR) is developed for rapid screening of plant-derived components in edible starch and its products. Methods Firstly, specific primers and probes were designed for peas, mung beans, cassava, and corn, and a multiplex real-time fluorescence PCR detection method was established. Furthermore, the specificity and absolute sensitivity of the method were verified, and finally, the limit of detection of the method was verified through adulteration simulation experiments. Results The method had strong specificity and couid only detect components derived from peas, mung beans, cassava and corn, the method had high sensitivity, the detection sensitivity for mung bean and cassava was 1×10–1 ng/μL, the detection sensitivity of pea and corn was 1×10–2 ng/μL. Conclusion The five fold real-time fluorescence PCR method established in this study can be used to distinguish pea derived, mung bean derived, corn derived, and cassava derived components in starch and its products on the market, and it has good practical value.