浙江农业学报

• 食品科学 • 上一篇    下一篇

利用环介导等温扩增技术检测茶叶中掺杂的大豆成分

  

  1. (宁波出入境检验检疫局,浙江 宁波 315012)
  • 出版日期:2015-08-25 发布日期:2015-08-21

Rapid detection of soybean mixture in tea by loop\|mediated isothermal amplification method#br#

  1. (Ningbo Entry\|Exit Inspection and Quarantine Bureau, Ningbo 315012, China)
  • Online:2015-08-25 Published:2015-08-21

摘要: 采用环介导等温扩增技术,建立了一种快速简便的检测茶叶中掺杂的大豆成分的方法。根据大豆内源基因序列,共设计了6条引物,通过环介导等温扩增技术(LAMP)扩增得到特征性的瀑布状条带。特异性检测表明,所设计的引物对大豆成分的检测具有良好的特异性;灵敏度试验显示,LAMP方法比荧光PCR方法灵敏度高10倍。通过对反应温度、甜菜碱浓度、Mg2+浓度和dNTP浓度的优化,本研究建立起优化的反应条件,整个检测周期约1 h,检测结果采用SYBR green I染色显示,易于观察和判定,可应用于大批量茶叶样本中掺杂大豆成分的检测。

关键词: 茶叶, 大豆, 环介导等温扩增, 检测

Abstract:  In the present study, a rapid and simple method to detect soybean mixture in tea by loop\|mediated isothermal amplification (LAMP) was established. On the basis of the endogenous gene lectin of soybean, six primers were designed and characteristic waterfall\|like bands by LAMP were acquired. Specificity tests showed that the primers were specific for soybean detection. Sensitivity test showed that LAMP was 10 times higher sensitive than fluorescence PCR. According to the optimized temperature, concentration of betaine, Mg2+, and dNTP, the entire testing cycle was about 1 h. Besides, it was convenient to observe the results by using SYBR green I. Thus, the established LAMP method could be useful for soybean component detection from tea samples in the near future.

Key words: tea, soybean, loop\, mediated isothermal amplification (LAMP), detection