多重实时荧光定量聚合酶链式反应在芝麻酱掺假鉴别中的应用

Application of multiplex real-time polymerase chain reaction for adulteration identification in sesame sauce

中文关键词:  多重实时荧光定量PCR  芝麻酱  植物源性成分  掺假鉴别

英文关键词:multiplex Real-time Polymerase Chain Reaction  sesame sauce  plant derived components  adulteration identify

中文摘要:

      目的 研究基于分子生物学技术, 建立多重实时荧光定量聚合酶链式反应(polymerase chain reaction, PCR)技术检测芝麻酱中的植物源性成分, 实现芝麻酱的快速掺假鉴别, 突破单标准单物种检测的局限性。方法 该研究以芝麻2S albumim mRNA基因、花生Ara b2基因、大豆Lectin基因、玉米adh1基因设计特异性引物和探针, 优化反应体系, 建立多重实时荧光定量PCR技术检测芝麻酱中的芝麻源性成分、花生源性成分、大豆源性成分、玉米源性成分, 并应用于市售的芝麻酱样本检测分析。结果 该方法高效低成本, 特异性好, 对小米、绿豆等10种非目标源性成分无特异性扩增; 对芝麻源性成分、花生源性成分、大豆源性成分、玉米源性成分的最低检出限为100 pg/μL, 具有较高的灵敏度; 应用建立的方法, 对市售21份芝麻酱样本进行检测分析, 检测结果与标签标注不符合的有4份, 标签标注符合率为80.95%, 与参比方法检测结果一致。

英文摘要:

      Objective To Research based on molecular biology technology, a multiplex real-time polymerase chain reaction technology was established to detect plant derived components in sesame sauce, achieving rapid adulteration identification of sesame sauce and breaking through the limitations of single standard single species detection. Methods In this study, specific primers and probes were designed with sesame 2S albumim mRNA gene, peanut Arab2 gene, soybean Lectin gene, and maize adh1 gene to optimize the reaction system, establish multiplex real-time PCR technology to detect the components of sesame, peanut, soybean, and corn in sesame paste, and apply it to the detection and analysis of sesame paste samples sold in the market. Results The results showed that this method was efficient, low-cost, and had good specificity. It did not specifically amplify 10 kinds of plant components such as millet and mung beans. The minimum detection limit for sesame, peanut, soybean and corn was 100 pg/ μL, with high sensitivity. 21 samples of sesame sauce from the market were tested and analyzed by using the established method. Four of the test results did not match the label labeling, with a label labeling compliance rate of 80.95%, which was consistent with the test results of the reference method. Conclusion The established multiplex real-time PCR technology has good applicability for the detection of processed products, providing a new molecular biology method for the identification of adulteration in sesame sauce.