基于实时荧光定量PCR检测特种乳中牛乳的掺伪

摘要/Abstract

摘要： 随着人们对特种乳需求的不断增加，特种乳制品中掺入低价值牛乳事件频发，乳制品的真实性越来越引起人们的重视。本研究分别基于SYBR Green实时荧光定量PCR和TaqMan实时荧光定量PCR技术检测特种乳及其热处理加工产品中掺假的牛乳成分，同时探讨了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。两种实时荧光定量PCR方法对于牛乳DNA的最低检测限分别为1 pg（SYBR Green）和10 pg（TaqMan），且均可以最低检测出特种乳混合物中0.1%（w/w）牛乳成分掺伪。为评估掺假模拟的重现性，在3天内共进行了9次重复试验，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。

关键词: 特种乳, Real-time PCR, 食品掺假, 真实性分析

Abstract: With the increasing demand for special milk, low value milk is often mixed into special dairy products, and the authenticity of dairy products attracts more and more attention. In this study, the adulterated milk components in special milk and its heat-treated products were detected based on SYBR Green quantitative real-time PCR and TaqMan quantitative real-time PCR, and the influence of different heat treatment methods on adulteration detection is discussed to meet the detection requirements of different commercial special dairy products. The results show that the designed bovine specific primers can amplify DNA in cow milk and have high specificity without cross-reactivity with non-target animals. The minimum detection limits of the two quantitative real-time PCR methods for milk DNA were 1 pg (SYBR Green) and 10 pg (TaqMan), respectively, and both of them could detect at least 0.1% (w/w) of milk adulteration in the special milk mixture. In order to evaluate the reproducibility of the adulterated simulation, a total of 9 repeated tests were carried out in 3 days to verify the high reproducibility of the method. In addition, the established quantitative real-time PCR method was used to analyze the authenticity of 20 commercial processed special dairy products. There are 7 products were detected to contain milk components, indicating deliberate adulteration. The experimental method is feasible for the authenticity detection of dairy products.

Key words: Special milk, Quantitative real-time PCR, Food adulteration, Authenticity analysis

中图分类号:

TS252.7

李威孙国栋鹿嘉榕赵仲凯杨洁. 基于实时荧光定量PCR检测特种乳中牛乳的掺伪[J]. 食品科学.

Wei LI Guo-Dong SUN Jie yang. Detection of adulteration of milk in special milk based on quantitative real-time PCR[J]. FOOD SCIENCE.

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