作　　者： ; ; (谭贵良）; (刘垚）; (张世伟）;

机构地区： 中山市食品药品检验所

出　　处： 《现代食品科技》 2015年第12期331-337,共7页

摘　　要： 建立了食用植物油和地沟油中动物源性成分的环介导等温扩增(LAMP)检测方法。针对猪、牛、鸡、鸭的线粒体Cytb基因分别设计2条外引物(F3、B3)、2条内引物(FIP、BIP)和2条环引物(FLP、BLP),着重对内引物FIP/BIP、甜菜碱和Mg2+浓度等反应参数进行了优化,扩增产物采用恒温实时荧光法检测。对方法的特异性和灵敏度进行了评价。实验结果表明,在0.2μmol/L外引物(F3和B3)、1.6μmol/L内引物(FIP和BIP)、0.8μmol/L环引物(FLP和BLP)、1 mol/L甜菜碱、6 mmol/L Mg SO4、1.6 mmol/L d NTP、反应温度63℃等参数的优化条件下,上述四种物源性成分的扩增效果最佳,该方法可以检测出动物油脂质量比例为1%及地沟油质量比例为5%的混合油脂中的动物源性成分。本方法对猪、牛、鸡、鸭源性成份的检测具有很高的特异性和灵敏度,可以用于油脂中动物源性成分的检测,为油脂的鉴伪和地沟油的鉴别提供技术支持。 A loop-mediated isothermal amplification (LAMP) method was established to detect the presences of animal-derived ingredients in edible vegetable oils and hogwash oils. Two outer primers (F3 and B3) and two inner primers (FIP and BIP) were designed based on the mitochondrial Cytb genes of pig, cattle, chicken, and duck. The inner primers FIP/BIP, betaine, Mg2+, and other reaction parameters were optimized, and the amplified products were detected with real-time fluorescence LAMP. The specificity and sensitivity of the method were evaluated. The results showed that the optimal conditions were 0.2 µmol/L outer primers (F3 and B3), 1.6 µmol/L inner primers (FIP and BIP), 0.8 μmol/L loop primers (FLP and BLP), 1 mol/L betaine, 6 mmol/L Mg2+, 1.6 mmol/L dNTPs, and a reaction temperature of 63. Under the optimal conditions, the best amplification was achieved for the abovementioned four animal-derived ingredients. With this improved LAMP approach, the animal-derived ingredients in mixed oils containing 1% of animal oils and 5% of hogwash oils could be detected. This method exhibits a very high specificity and sensitivity for pig-, cattle-, chicken-, and duck-derived ingredients, can be applied in the detection of animal-derived ingredients in oils, and provides a technical support for identification of oils and hogwash oils. © 2015, South China University of Technology. All right reserved.

关 键 词： 食用植物油 地沟油 环介导等温扩增 动物源性成分 基因 鉴别

领　　域： [轻工技术与工程] [轻工技术与工程]