基于实时聚合酶链式反应检测特种乳中牛乳的掺伪  

Detection of Bovine Milk Adulteration in Non-novine Milk Using Real-Time Polymerase Chain Reaction

作　　者：李威 孙国栋 鹿嘉榕 赵仲凯 杨洁 LI Wei;SUN Guodong;LU Jiarong;ZHAO Zhongkai;YANG Jie(Xinjiang Key Laboratory of Biological Resources and Genetic Engineering,College of Life Science and Technology,Xinjiang University,Ürümqi 830017,China;Xinjiang Camel Industry Engineering and Technology Research Center,Ürümqi 830017,China)

机构地区：[1]新疆大学生命科学与技术学院,新疆生物资源基因工程重点实验室,新疆乌鲁木齐830017 [2]新疆骆驼产业工程技术研究中心,新疆乌鲁木齐830017

出　　处：《食品科学》2025年第6期263-274,共12页Food Science

基　　金：新疆维吾尔自治区自然科学基金项目(2022D01C680);新疆维吾尔自治区重点研发专项(2023B02034);新疆维吾尔自治区创新环境(人才、基地)建设专项(20200061)。

摘　　要：分别基于SYBR Green实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)和Taq Manreal-timePCR技术检测特种乳及其热处理加工产品中掺假的牛乳成分,同时探讨不同热处理方式对掺假检测的影响,以满足不同商品化特种乳制品的检测要求。结果表明,所设计的牛特异性引物可以扩增牛乳中的DNA,与非目标动物无交叉反应性,具有较高的特异性。两种real-time PCR方法对于牛乳DNA的检测限分别为1 pg(SYBR Green)和10 pg(Taq Man),且均可以最低检测出特种乳混合物中0.1%(m/m)牛乳成分掺伪。为评估掺假模拟的重现性,在3 d内共进行了9次重复实验,验证了方法具有较高的重复性。同时,为检测本研究中所建立实验方法的可用性,对20种商业加工特种乳制品进行了真实性分析,其中在7种产品中检测出牛乳成分,显示出所建立的检测方法具有较高的分辨率和实际应用价值。In this study,adulteration of bovine milk in fresh and thermally processed non-bovine milk was detected by SYBR Green and TaqMan real-time polymerase chain reaction(real-time PCR),and the effect of different heat treatment methods on the adulteration detection was explored aiming to meet the requirements for the detection of adulteration of different commercial non-bovine milk products.The results showed that the designed bovine-specific primers allowed specific amplification of DNA in bovine milk without cross-reactivity with DNA from non-target animals.The minimum detection limits of SYBR Green and TaqMan real-time PCR for bovine milk DNA were 1 and 10 pg,respectively,and both methods were capable of detecting at least 0.1%(m/m)bovine milk adulteration in non-bovine milk mixtures.The results of nine repeated experiments over three days confirmed high reproducibility of the methods.In addition,using the real-time PCR methods,bovine milk was detected in 7 out of 20 commercial non-bovine milk products,indicating high resolution and great practical application value of these methods.

关 键 词：特种乳 实时聚合酶链式反应 食品掺假 真实性分析 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]