利用环介导等温扩增技术检测茶叶中掺杂的大豆成分  

Rapid detection of soybean mixture in tea by loop-mediated isothermal amplification method

作　　者：余澍琼[1] 张吉红[1] 崔俊霞[1] 张慧丽[1] 倪健波[1] 

机构地区：[1]宁波出入境检验检疫局,浙江宁波315012

出　　处：《浙江农业学报》2015年第8期1479-1483,共5页Acta Agriculturae Zhejiangensis

基　　金：宁波市科研项目(2011C50080);宁波检验检疫局项目(甬K07-2011);质检公益性行业科研专项项目(201010256);浙江省重点科技创新团队(2010R50028)

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d NTP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。In the present study, a rapid and simple method to detect soybean mixture in tea by loop-mediated isother- mal 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 fluores- cence PCR. According to the optimized temperature, concentration of betaine, Mg2＋ , and dNTP, the entire testing eycle 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.

关 键 词：茶叶 大豆 环介导等温扩增 检测 

分 类 号：S571.1[农业科学—茶叶生产加工] Q819[农业科学—作物学]