Rapid authentication of the precious herb saffron by loop-mediated isothermal amplification (LAMP) based on internal transcribed spacer 2 (ITS2) sequence.
Rapid authentication of the precious herb saffron by loop-mediated isothermal amplification (LAMP) based on internal transcribed spacer 2 (ITS2) sequence.
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DOI：
10.1038/srep25370
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发表时间：
2016-05-05
影响因子：
4.6
通讯作者：
Chen S
中科院分区：
文献类型：
--
作者：
Zhao M;Shi Y;Wu L;Guo L;Liu W;Xiong C;Yan S;Sun W;Chen S
藏红花是最昂贵的中药材之一，由于价格昂贵且产量有限，一直受到各种掺假的影响。本研究介绍了一种环介导等温扩增 (LAMP) 技术，用于区分藏红花及其掺假品。这种新颖的技术灵敏、高效且简单。根据藏红花内转录间隔区2（ITS2）核糖体DNA的核苷酸序列，设计了6种特异性LAMP引物。所有 LAMP 扩增均成功进行，在 65°C 等温条件下，目视检测在 60 分钟内完成。结果表明，LAMP 引物对于区分藏红花及其掺假品是准确且高度特异的。特别是，10fg 基因组 DNA 被确定为藏红花中 LAMP 模板准确性的极限。因此，所提出的新颖、简单且灵敏的 LAMP 测定非常适合草药材料的即时现场辨别。基于该研究，提供了利用 LAMP 协议进行草药认证的实用标准操作程序（SOP）。
Saffron is one of the most expensive species of Chinese herbs and has been subjected to various types of adulteration because of its high price and limited production. The present study introduces a loop-mediated isothermal amplification (LAMP) technique for the differentiation of saffron from its adulterants. This novel technique is sensitive, efficient and simple. Six specific LAMP primers were designed on the basis of the nucleotide sequence of the internal transcribed spacer 2 (ITS2) nuclear ribosomal DNA of Crocus sativus. All LAMP amplifications were performed successfully, and visual detection occurred within 60 min at isothermal conditions of 65 °C. The results indicated that the LAMP primers are accurate and highly specific for the discrimination of saffron from its adulterants. In particular, 10 fg of genomic DNA was determined to be the limit for template accuracy of LAMP in saffron. Thus, the proposed novel, simple, and sensitive LAMP assay is well suited for immediate on-site discrimination of herbal materials. Based on the study, a practical standard operating procedure (SOP) for utilizing the LAMP protocol for herbal authentication is provided.