实时荧光PCR法在鱼翅鉴别中的应用

摘要

为有效鉴别市售鱼翅的真伪,针对鱼翅中鲨鱼源性成分的线粒体基因组的部分序列,利用实时荧光聚合酶链式反应(Polymerase Chain Reaction,PCR)技术,通过特异性引物和探针的设计建立起一种有效的真伪鉴别方法.该方法中对样品脱氧核糖核酸(Deoxyribonucleic acid,DNA)的提取方法、DNA检测的浓度灵敏度、质量灵敏度以及方法的特异性进行研究.结果表明鱼翅样品用试剂盒进行提取效果良好,其检测的DNA浓度灵敏度可达1×10-5 ng/μL,质量灵敏度可达0.01%.利用该方法对市场上的购买的47份鱼翅类样品和18份非鱼翅样品进行了检测,43样品检测出鲨鱼成分,包括4份仿鱼翅在内的22份未检出鲨鱼成分.%A novel effective method for identification of shark fins available in market was established by Real time Polymerase Chain Reaction (PCR) technique. Specific primers and probes were designed from a portion mitochondria genome of shark fin. Extraction method of Deoxyribonucleic acid(DNA) , sensitivity of DNA con-centration, sensitivity of DNA weight and specificity were investigated. It was indicated that enough concentra-tion of DNA for detection would be obtained by kit extraction method. DNA concentration sensitivity of 1 × 10-5 ng/μL and DNA weight sensitivity of 0.01%were abtained. 47 shark fin products and 18 non-shark fin products available in market were determined with the above PCR method. DNA of shark was be found in 43 products while DNA of shark was not be found in other 22 products including 4 man-made shark fin products.