实时荧光PCR技术快速检测莲蓉制品中芸豆成分

发布时间：2018-07-20 15:51

【摘要】：目的:建立基于实时荧光聚合酶链式反应(polymerase chain reaction,PCR)技术检测莲蓉制品中芸豆成分的方法。方法:以芸豆pvsbe2基因高度保守区域设计特异性引物和探针,通过对莲子及其他富含淀粉类植物DNA进行扩增,以验证方法的特异性;以1 ng/μL的芸豆DNA进行系列稀释,确定此法检测灵敏度;对含有1%芸豆与莲子的混合样品的DNA模板进行10倍梯度稀释,确定重量检测灵敏度;并应用此方法和PCR方法对市场样品进行了检测,对建立的荧光PCR方法进行验证。结果:该检测方法具有高度特异性,与莲子及其他高淀粉植物无交叉反应;DNA质量浓度的检测灵敏度达到1 pg/μL,重量检测灵敏度可达0.01%。含芸豆成分的莲蓉月饼扩增阳性,检测结果与食品标签相符。结论:本研究建立的实时荧光PCR方法具有特异性强、灵敏度高、快速简便的特点,更适合莲蓉制品中芸豆成分的快速检测。
[Abstract]:Aim: to establish a real-time fluorescent polymerase chain reaction (polymerase chain) method for the determination of kidney bean in lotus paste products. Methods: specific primers and probes were designed with highly conserved region of pvsbe2 gene of kidney bean to amplify lotus seed and other starch-rich plants to verify the specificity of the method. The sensitivity of this method was determined. The DNA template containing 1% kidney bean and lotus seed was diluted by 10 times gradient to determine the sensitivity of gravimetric detection. The established fluorescent PCR method was verified. Results: this method has a high specificity. The sensitivity of the method is 1 pg/ 渭 L without cross reaction with lotus seed and other high starch plants, and the sensitivity of weight detection is 0. 01%. Lotus paste mooncakes containing kidney bean components were amplified positive, and the results were consistent with food labels. Conclusion: the real-time PCR method developed in this study has the characteristics of high specificity, high sensitivity, rapid and simple, and is more suitable for rapid detection of kidney bean components in lotus seed paste products.
【作者单位】：广西梧州食品药品检验所;
【分类号】：O657.3;TS207.3

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