Rapid visual detection of phytase gene in genetically modified maize using loop-mediated isothermal amplification method
Rapid visual detection of phytase gene in genetically modified maize using loop-mediated isothermal amplification method
复制标题
DOI：
10.1016/j.foodchem.2014.01.102
复制
发表时间：
2014-08-01
期刊：
影响因子：
8.8
通讯作者：
Chen, Hongjun
中科院分区：
文献类型：
--
作者：
Huang, Xin;Chen, Lili;Chen, Hongjun
2009年我国已报道并批准了转植酸酶基因玉米。本文报道了一种高特异性的环介导等温扩增（LAMP）方法检测转基因玉米中的植酸酶基因。LAMP反应需要不到20分钟，扩增是可见的，无需凝胶电泳。LAMP方法检测植酸酶基因组DNA的灵敏度约为30个拷贝，是常规PCR凝胶电泳法的33.3倍。定量检测结果表明，在6 × 10 ~（1）~ 6 × 107拷贝的动态范围内，LAMP方法的Tt值与DNA拷贝数呈良好的线性关系，定量限为60拷贝。因此，与传统PCR技术相比，LAMP方法直观、快速、灵敏，不需要特殊设备，对于转基因饲料的现场测试和快速筛查非常有用。(C)2014爱思唯尔有限公司版权所有。
Transgenic maize plant expressing high phytase activity has been reported and approved by Chinese government in 2009. Here, we report a highly specific loop-mediated isothermal amplification (LAMP) method to detect the phytase gene in the GMO maize. The LAMP reaction takes less than 20 min and the amplification is visible without gel electrophoresis. The detection sensitivity of the LAMP method is about 30 copies of phytase genomic DNA, which is 33.3 times greater than the conventional PCR method with gel electrophoresis. The quantitative detection results showed that the LAMP method has a good linear correlation between the DNA copy number and the associated Tt values over a large dynamic range of template concentration from 6 x 10(1) to 6 x 107 copies, with a quantification limit of 60 copies. Therefore, the LAMP method is visual, faster, and more sensitive, and does not need special equipment compared to traditional PCR technique, which is very useful for field tests and fast screening of GMO feeds. (C) 2014 Elsevier Ltd. All rights reserved.