水牛乳及其制品中掺入奶牛乳的单重和双重PCR检测方法  

Research of detecting adulteration of dairy cow milk into buffalo milk and its products by simplex and duplex PCR method

作　　者：林波[1] 黎颖 李玲[1] 杜寒春 唐艳[1] 农皓如[1] 曾庆坤[1] 

机构地区：[1]中国农业科学院广西水牛究所广西水牛遗传繁育重点实验室,南宁530001 [2]广西分析测试中心,南宁530022

出　　处：《中国乳品工业》2013年第6期35-38,共4页China Dairy Industry

基　　金：广西公益性科研院所基本科研业务费(2012ACZ06);广西科学研究与技术开发项目(No.12118011-2A)

摘　　要：应用奶牛特异性引物,采用苯酚-氯仿抽提法提取水牛乳DNA后,分别以单重和双重PCR法对掺入荷斯坦牛乳量为0%,0.5%,5%,20%,50%和100%的新鲜水牛奶、高温杀菌水牛奶、水牛乳酸奶和水牛乳Mozzarella干酪进行了检测。结果表明,单重PCR方法最低可检出新鲜水牛奶、水牛乳Mozzarella干酪和水牛乳酸奶中掺入0.5%的荷斯坦牛乳成分,而对高温杀菌水牛奶则最低只能检出5%的掺入比例;以相同引物进行双重PCR检测的结果与单重PCR结果一致,可实现PCR检测一步完成。因此,PCR法可用于水牛乳及其乳制品中掺入荷斯坦牛乳的鉴别,特别是双重PCR法可经一步PCR完成检测,具有方便和快速的特点。This study applies the cow specificity primer, using the chloroform/phenol method to extract DNA from buffalo milk, then the simplex and duplex method PCR method were applied to detect adulteration of Holstein cow into fresh buffalo milk, high temperature steril- ized buffalo milk, buffalo yogurt and Mozzarella cheese at level of 0%, 0.5%, 5%, 20%, 50% and 100%, respectively. The results showed that the simplex PCR method successfully detected the adulteration level as low as 0.5% in fresh milk, mozzarella cheese and yogurt, while the adulteration level only can be detected by 5% in high temperature sterilized buffalo milk. The results of duplex PCR assay was the same as simplex PCR results, while the detection can be finished in only one step PCR. Therefore, PCR can be used to identify the adulteration of Holstein cow milk into buffalo milk or its products, especially, the duplex PCR method can finish the assay in one step, and has the advantages of fast and convenient.

关 键 词：水牛乳 奶牛乳 乳制品 掺假 PCR 检测方法 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]