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目的建立可同时检测掺入羊肉中的猪、马、牛、鸭成分的多重荧光PCR检测体系。方法配制掺入猪、马、牛、鸭源性成分的模拟羊肉掺假样品,掺假比例17%~45%。筛选特异性强、敏感度高的猪、马、牛、鸭物种特异性基因,优化反应体系,建立了针对猪、牛、马、鸭两两配对的两重荧光PCR检测体系和其中3个任意组合的三重荧光PCR检测体系。结果模拟羊肉掺假样品两重荧光PCR检测结果显示,包括单个成分掺入比例低至7%的样品,其检测准确率为100%。而三重荧光PCR检测同时检测猪、牛、马、鸭四种肉品中的3种,其中猪肉、牛肉和马肉的掺假比例均为8%~15%;鸭牛肉掺假比例为5%~10%。三重荧光PCR检测结果显示,所有掺假样品中的各种掺假组分都被准确检出。结论本研究建立的多重荧光PCR检测体系能准确检出配制的模拟掺假羊肉制品中的猪、马、牛、鸭成分,可进一步研究以应用于市场肉制品及加工肉制品掺假的检测。
Objective To establish a multiplex fluorescence PCR detection system that can simultaneously detect the composition of pigs, horses, cattle and ducks incorporated into mutton. Methods Adulterated mutton sample of imitation mutton mixed with pigs, horses, cattle and ducks, with adulteration ratio of 17% ~ 45%. The specific genes with high specificity and high sensitivity were screened for the specific gene of porcine, horse, cattle and duck and the reaction system was optimized. A double fluorescent PCR detection system was established for pairs of pigs, cattle, horses and ducks and three of them Combined triplex fluorescence PCR detection system. Results The results of two-fold fluorescent PCR assay of simulated adulterated mutton samples showed that the detection accuracy was 100% for the samples with a single ingredient incorporated in a proportion of as low as 7%. The triple fluorescent PCR detection simultaneously detected three kinds of pork, beef, horse and duck four kinds of meat products, including pork, beef and horse meat adulteration rates were 8% to 15%; duck beef adulteration rate was 5% ~ 10%. Triple fluorescence PCR results showed that all adulterated components in all adulterated samples were accurately detected. Conclusion The multiplex fluorescence PCR detection system established in this study can accurately detect the composition of pigs, horses, cattle and ducks in simulated adulterated mutton products and can be further studied for the detection of adulterated flesh products and processed meat products.