牛羊乳区别检验的PCR高分辨熔解检测方法

Identification of goat,sheep,bovine and water buffalo origin component in dairy products using high-resolution melting-PCR

乳源成分鉴别在乳制品掺假检测中具有重要意义。为了检测乳及其制品中牛、水牛、山羊、绵羊4种动物源性成分,建立了基于通用引物的PCR高分辨熔解曲线(high-resolution melting,HRM)检测方法。通过分析通用引物扩增子的熔解曲线,鉴别了含多种动物源性成分的混合物,检出了在山羊乳、水牛乳中掺入5%的牛乳,并准确判别市售乳及乳制品的乳源成分。该方法快速、灵敏、通量高,可以实现1对引物区分牛、水牛、山羊、绵羊4种动物源性成分,是一种有效的乳源成分鉴别方法,也可以用于检测肉及肉制品等其他含有4种以上动物源性成分的食品。

The identification of the species origin is of great significance in dairy products adulteration detection.In order to detect four common animal species(bovine,buffalo,goat and sheep)in milk and dairy products,a PCR high-resolution melting(HRM)detection method was established based on universal primers detection.By analyzing the melting curve of the universal primer-generated amplicons,PCR-HRM was capable of identifying the mixtures of various animal-derived components,and also of sensitively detecting the presence of bovine milk in goat milk and buffalo milk down to 5%.In addition,it could accurately identify the species origin of commercial milk and dairy products.The proposed method is rapid,sensitive,high throughput,and can realize the identification of bovine,buffalo,goat and sheep with only one pair of primer.It is an effective method for the identification of milk derived constituents.It could be further applied to identify the above four animal species in various meat and meat products or other foods.