Reduced Graphene Oxide-Based Solid-Phase Extraction for the Enrichment and Detection of microRNA
Reduced Graphene Oxide-Based Solid-Phase Extraction for the Enrichment and Detection of microRNA
复制标题
基于还原氧化石墨烯的固相萃取用于 microRNA 的富集和检测
DOI：
10.1021/acs.analchem.7b03138
复制
发表时间：
2017-10-03
影响因子：
7.4
通讯作者：
Xing, Wanli
中科院分区：
文献类型：
--
作者：
Yan, He;Xu, Youchun;Xing, Wanli
MicroRNA (miRNA) 是具有调节功能的内源性分子。 miRNA的纯化和富集对于其精确和灵敏的检测至关重要。使用商业试剂盒分离的miRNA含有丰富的干扰RNA，并且miRNA的浓度可能不足以检测。在此，我们制备了一种基于还原氧化石墨烯（rGO）的磁性固相萃取材料，用于从复杂的核酸溶液中富集和超灵敏检测 miRNA。原位逆转录 (RT) 被开发为迄今为止报道的与后续扩增兼容的方法中从 rGO 解吸 miRNA 的最有效方法。此外，使用滚环扩增和qPCR检测let-7a，检测限分别降低了24.7倍和31.3倍。该材料还成功用于从人血浆中分离的总RNA中提取和检测miRNA。我们的结果表明，在科学研究中临床发现新的 miRNA 的材料。
MicroRNAs (miRNAs) are endogenous molecules with regulatory functions. The purification and enrichment of miRNA are essential for its precise and sensitive detection. miRNA isolated using commercial kits contains abundant interfering RNAs, and the concentration of miRNA may not be adequate for detection. Herein, we prepared a reduced graphene oxide (rGO)-based magnetic solid-phase extraction material for the enrichment and ultrasensitive detection of miRNA from intricate nucleic acid solutions. In situ reverse transcription (RT) was developed as the most efficient approach to desorb miRNA from rGO among the methods that are compatible for the subsequent amplification reported thus far. Additionally, rolling circle amplification and qPCR were used to detect let-7a with a decrease of the limit of detection by 24.7- and 31.3-fold, respectively. This material was also successfully used to extract and detect miRNA from total RNA isolated from human plasma. Our results show that the material in clinics discovery of new miRNAs in scientific research.