PCR法检测鱼及其制品中的鱼源性成分

摘要

Objective:To establish a rapid,specific and sensitive polymerase chain reaction (PCR) method for the detection of fish-derived ingredients.Methods:A set of primers was used to selectively amplify a short DNA fragment of the fish mitochondrial 12S rRNA gene.The specificity was evaluated by analyzing 24 kinds of fish as well as meat,poultry and shrimp,commonly added in fish products.The sensitivity was evaluated by amplifying DNA extracted from grass carp mixed with meat from other animal species.Results:The method established could specifically detect fish-derived ingredients with a sensitivity of 0.5％.Conclusion:This PCR method is a rapid,sensitive and specific method for preliminary screening of fish-derived ingredients in foods,and is of important significance to safeguard the interests of consumers and standardize the market order.%目的:建立一种快速、特异、灵敏的鱼源性成分聚合酶链式反应(polymerase chain reaction,PCR)检测方法.方法:根据鱼线粒体基因组12S rRNA中的保守序列设计鱼源性特异性引物,进行PCR扩增,建立鱼源性成分检测方法;对24种鱼及鸡、牛、羊、猪、鸭、虾6种常见的易混于鱼制品的动物源性成分进行特异性检测;将草鱼肉混入其他动物肉中,混合均匀后提取DNA进行PCR扩增,确定肉样水平的检测灵敏度;将草鱼DNA混入其他动物DNA中,以混合后的DNA为模板进行PCR扩增,确定DNA水平的检测灵敏度.结果:该方法能特异性的对鱼源性成分进行快速检测,检测灵敏度达0.5％.结论:该方法能对食品中是否含有鱼源性成分进行初筛,到达快速检测的目的,对防止食品掺假、维护消费者利益、规范市场秩序有重要意义.