实时荧光定量PCR法鉴别驴乳中的牛乳、羊乳成分

Identification of Bovine and Goat Milk Components in Donkey Milk by Real-Time Fluorescence Quantitative PCR

建立了一种实时荧光定量PCR法检测驴乳中牛源性、羊源性的检测方法。将牛乳、羊乳分别以0.05%、0.1%、0.2%、0.3%、0.4%、0.5%、0.6%、0.7%、0.8%、0.9%和1.0% (v/v)的比例添加进驴乳中,制成牛乳和驴乳、羊乳和驴乳的混合乳样品。提取DNA后用实时荧光定量PCR仪检测。结果表明:驴乳中掺杂牛乳、羊乳后可鉴别出,检出限分别为0.3%、0.1%。该方法通量大、灵敏度高、特异性好,实现了对驴乳中牛、羊源性成分快速、准确的检测,对保障相关产品市场安全具有重要意义。

A real-time fluorescence quantitative PCR methodwas established for the determination of bovine and sheep origin in donkeymilk. The mixed milk samples of cow milk and donkey milk, goat milk and donkeymilk were prepared by adding cow milk and goat milk into donkey milk at theproportions of 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% and1.0% (V/V), respectively. DNA was extracted and detected by real-time fluorescencequantitative PCR. The results showed that donkey milk adulterated with cow’smilk and goat’s milk could be identified and the detection limits were 0.3% and0.1%, respectively. This method has high throughput, high sensitivity and good specificity,realizing the rapid and accurate determination of bovine and sheep derived componentsin donkey milk. It had great significance for ensuring the security of related products.