食品科学 ›› 2009, Vol. 30 ›› Issue (12): 190-193.doi: 10.7506/spkx1002-6630-200912041

• 分析检测 • 上一篇    下一篇

实时荧光P C R 在鉴别奶粉中掺入大豆成分的应用研究

岳巧云,陈定虎,伍朝晖,周灼标,邱德义*   

  1. 中山出入境检验检疫局技术中心
  • 收稿日期:2008-08-23 修回日期:2008-11-19 出版日期:2009-06-15 发布日期:2010-12-29
  • 通讯作者: 邱德义 E-mail:qiudy@zs.gdciq.gov.cn, dqiu.madrid@gmail.com

Real-Time Fluorescent PCR Detection of Soybean Ingredients Adulterate in Milk Powder

YUE Qiao-yun,CHEN Ding-hu,WU Zhao-hui,ZHOU Zhuo-biao,QIU De-yi*   

  1. (Centre of Inspection and Quarantine Technology, Zhongshan Entry-Exit Inspection and Quarantine
    Bureau, Zhongshan 528403, China)
  • Received:2008-08-23 Revised:2008-11-19 Online:2009-06-15 Published:2010-12-29
  • Contact: QIU De-yi* E-mail:qiudy@zs.gdciq.gov.cn, dqiu.madrid@gmail.com

摘要:

首次利用实时荧光PCR 方法,以大豆内源参照基因lectin 为指标,对奶粉中掺入豆粉等植物成分进行定性、定量分析研究。结果表明,通过正己烷抽提奶粉中的油脂,苯酚抽提奶粉中的蛋白质等处理步骤后,应用植物基因组DNA 提取试剂盒能够得到高质量的DNA 模板。实时荧光PCR 反应结果表明,该方法判定奶粉中掺入植物成分的灵敏度大大提高;奶粉中掺入大豆粉或大豆制品的掺入量与实时荧光PCR扩增曲线及循环数呈良好的对应关系。实时荧光PCR 法能够对奶粉中掺入的植物成分进行快速准确地定性、定量分析,可以作为市场监督和检验鉴定的可行性方法。

关键词: 奶粉, 实时荧光PCR, 植物蛋白成分, DNA 提取纯化

Abstract:

Soybean ingredients adulterated in milk powder was firstly determined by real-time Fluorescent PCR in our study using lectin, the endogenous gene of soy-bean as a reference gene. This method provided a quick and precise way for the detection of adulteration of milk powder and also presented new insights for the market supervision and administration. After respective removal of fat and protein in milk powder with hexane and tris-balanced phenol, a high-quality DNA template was obtained using Promega Wizard genomic DNA purification kit. The experimental results indicated that the sensitivity of detecting soybean ingredients adulterated in milk powder was highly improved by using real-time fluorescent PCR method, and the PCR amplification curve and cycle numbers remained corresponding relationship with adulteration amounts of soybean ingredients in milk powder. This method can provide a rapid and accurate way for detection and analysis of the adulteration of milk powder, which can be used as a new evidence for market supervision and administration.

Key words: milk powder, real-time fluorescent PCR, plant protein ingredients, DNA extraction and purification

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