Rapid and Sensitive Detection of sFAT-1 Transgenic Pigs by Visual Loop-Mediated Isothermal Amplification
Rapid and Sensitive Detection of sFAT-1 Transgenic Pigs by Visual Loop-Mediated Isothermal Amplification
复制标题
通过视觉环介导的等温扩增快速、灵敏地检测 sFAT-1 转基因猪。
DOI：
10.1007/s12010-016-2041-8
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发表时间：
2016-07-01
影响因子：
3
通讯作者：
Zhou, Rong
中科院分区：
文献类型：
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作者：
Tao, Chenyu;Yang, Yalan;Zhou, Rong
转基因家畜消耗更少的资源和减少废物产生，有可能为改善环境和人类健康做出贡献。然而，转基因过程也带来了风险。转基因动物产品的安全性评价和控制已引起广泛关注，相关法规和技术正在制定中。快速检测技术在现场和海关抽样中发挥着重要作用。目前，环介导的等温扩增(LAMP)技术以其简便、快速、高效、特异等优点被广泛应用于核酸分析中。本研究设计了一种特异、灵敏的检测SFAT-1转基因猪的检测系统。针对该靶序列设计了一套包括两个环引物在内的六对引物。在优化的温度下，DNA样品在不到1h的时间内被扩增，并用LA-320C浊度计和直接添加钙黄绿素的肉眼检测。SFAT-1 LAMP的检测下限为1.26 ng/mU L，并对转基因和非转基因DNA样品进行了盲法检测。因此，本研究的结果表明，LAMP是一种非常有用的转基因检测工具。
Genetically modified (GM) livestock have the potential to contribute to improving the environment and human health, with consumption of fewer resources and reduced waste production. However, the transgene process also poses risks. The safety assessment and control of transgenic animal products have drawn wide attention, and the relevant regulations and technology are being developed. Quick testing technology plays a significant role in on-site and customs sampling. Nowadays, loop-mediated isothermal amplification (LAMP) was widely applied in nucleic acid analysis because of its simplicity, rapidity, high efficiency and specificity. In this study, a specific, sensitive detection system for detecting sFAT-1 transgenic pigs was designed. A set of six primers including two loop primers was designed for the target sequence. The DNA samples were amplified in less than 1 h at the optimized temperature and detecting by both Nephelometer LA-320c and unaided eyes directly adding calcein. The detection limit of sFAT-1 LAMP was as low as 1.26 ng/mu L. Furthermore, blind tests of transgenic and non-transgenic DNA samples were all correctly detected. Hence, the results in this study demonstrated that LAMP is a very useful tool for transgenic detection.