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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。
In this paper, the detection and identification of sharks in shark fin has developed a rapid and sensitive PCR detection method for the detection of shark fin food ingredients. According to the shark mitochondrial cytochrome subunit I gene sequence designed shark-specific primers, the length of 228 bp; To evaluate the specificity of the method, the designed primers for 22 shark fin sample DNA and 37 other kinds of DNA were PCR results showed that only a specific 228 bp band was detected in shark shark fin and no band was detected in the other 37 species. In order to evaluate the sensitivity of the method, PCR analysis of samples containing different proportions of potato DNA in shark’s fin DNA using this method revealed a detectable sensitivity of 0.1% (m / m). Forty-five different shark-fin samples were randomly selected and shark’s fin was found in 22 sharks, while 21 shark’s fin did not contain sharks but contained plant ingredients. The sample pretreatment method, the DNA extraction method and the PCR detection method can be widely applied to the detection and identification of shark components in food.