鱼翅类食品中鲨鱼成分PCR鉴定方法研究

Identification of Shark Fins in Food with PCR Method

摘要:

本文针对鱼翅中的鲨鱼成分进行检测鉴定开发了一种快速灵敏的PCR检测方法，可检测鱼翅类食品中是否存在鲨鱼成分。根据鲨鱼线粒体的细胞色素亚基I基因序列设计了鲨鱼特异性引物，扩增长度为228 bp；为了评价方法的特异性，将设计的引物分别针对22份鱼翅样品DNA和37种其它种类DNA进行PCR检测，结果显示，只在鲨鱼鱼翅中能检测出特异的228 bp条带，其它37种物种中均无条带检出。为了评价方法的灵敏度，将鱼翅DNA中掺入了不同比例土豆DNA的样品采用本方法进行了PCR分析，显示方法可检测灵敏度为0.1%（m/m）。随机抽取45份不同类型的鱼翅样品，检测出22份鲨鱼翅均含鲨鱼成分，而21份仿鱼翅均不含鲨鱼成分而含有植物成分。该样品前处理方法、DNA提取方法以及PCR 检测方法可广泛应用于食品中鲨鱼成分检测鉴定。

Abstract:

In order to detect the shark ingredients in food, a rapid and sensitive polymerase chain reaction (PCR) method was developed. In this method, a pair of primers was designed based on cytochrome oxidase subunit I (COI) gene, which could produce 228bp band via PCR. To evaluate the specificity of this method, 22 shark fin samples DNA and 37 other species DNA were analyzed by PCR using the new specific primers. The results showed that the specific 228 bp band only be observed in 22 shark fin samples. The sensitivity of the PCR method was tested by mixing shark fin DNA and DNA extracted from potato in different proportions, and it was indicated that the lowest detection limit by this method was 0.1% (m/m). Forty-three samples were randomly tested, 22 of them containing shark fins, while 21 imitation samples were detected to contain plant ingredients. This method can be widely used in identification of shark ingredients in food.