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肉类掺假不仅损害消费者利益,而且易引发民族矛盾,甚至会让过敏人群产生严重的过敏反应,因此对市场上销售的肉的种类进行快速、有效的鉴别意义重大。利用SYBRGreen实时PCR的方法,采用对家禽和哺乳动物Myostatin基因都有特异性扩增的通用引物MY,以确保所提取肉类DNA的质量。利用SYBRGreen实时PCR的溶解曲线对肉类成分进行检测判断。结果表明,在SYBRGreen实时PCR系统中,MY通用引物能够对市场上常见肉的DNA(猪、牛、山羊、绵羊、鸡、鸭)进行扩增。在利用猪肉冒充羊肉的检测中,能够利用两对引物的平均溶解温度的不同[猪Tm=(82.03±0.25)℃;羊Tm=(78.83±0.16)℃]对猪、羊肉进行鉴别,能够从1%猪肉+99%羊肉的混合物中检测出1%猪肉(0.1ng),从0.1%羊肉+99.9%猪肉的混合物中检测出0.1%(0.01ng)羊肉。
Meat adulteration not only harm the interests of consumers, but also easily lead to ethnic conflicts, and even make allergic people have a serious allergic reaction, so the types of meat on the market for rapid and effective identification of great significance. Using SYBRGreen real-time PCR, a universal primer, MY, was used to amplify the Myostatin gene in poultry and mammals to ensure the quality of extracted meat DNA. Using SYBRGreen real-time PCR dissolution curve to detect meat ingredients to determine. The results showed that the MY universal primers were able to amplify the common meat DNA (pigs, cattle, goats, sheep, chickens, ducks) on the market in the SYBR Green real-time PCR system. Pigs and mutton can be identified by using the difference between the average solubilization temperatures of two pairs of primers [pig Tm = (82.03 ± 0.25) ° C; sheep Tm = (78.83 ± 0.16) ° C] 1% pork (0.1 ng) was detected in a mixture of 1% pork + 99% lamb and 0.1% (0.01 ng) of lamb from a mixture of 0.1% lamb + 99.9% pork.