Highly sensitive and multiplexed miRNA analysis based on digitally encoded silica microparticles coupled with RCA-based cascade amplification
Highly sensitive and multiplexed miRNA analysis based on digitally encoded silica microparticles coupled with RCA-based cascade amplification
复制标题
基于数字编码二氧化硅微粒和基于 RCA 的级联扩增的高灵敏度和多重 miRNA 分析
DOI：
10.1039/c8an01393d
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发表时间：
2018-11-07
期刊：
影响因子：
4.2
通讯作者：
Li, Jiong
中科院分区：
文献类型：
--
作者：
Liu, Shengquan;Fang, Han;Li, Jiong
目前，miRNA分析对于理解miRNA调控网络和临床诊断治疗具有重要意义。对于多重miRNA敏感性分析可行的分析策略仍然有很高的需求。在此，我们提出了一种新的策略，通过耦合级联扩增与数字编码的二氧化硅微粒的miRNA分析。通过一步沉积策略以数字形式精确制造微粒，并且对于多重分析是高效的。由RCA和切口辅助链置换扩增（SDA）组成的级联扩增表现出高扩增效率，并且不需要复杂的序列设计，从而提高了与我们的微粒上的碱基堆积杂交的兼容性。已经实现了let-7a和miR-21在一锅中的平行和灵敏分析，而没有相互干扰，具有高灵敏度（LOD，约0.5 fM）和宽动态范围（10 pM-1 fM）。此外，我们的策略表现出高特异性的同源序列的miRNA和良好的抗干扰能力，在复杂的样品基质。考虑到有多达128（2（7））种微粒可用，我们的策略可以在一锅中应用于数十种miRNA敏感性分析，并且它在miRNA签名分析以及广泛的临床应用中具有巨大的潜力。
Currently, miRNA analysis is significant for understanding miRNA regulation networks and clinical diagnostics and therapy. Analytical strategies feasible for multiplex miRNA-sensitive analysis are still in high demand. Herein, we propose a novel strategy for miRNA analysis by coupling cascade amplification with digitally encoded silica microparticles. The microparticles are precisely fabricated in a digital form through a one-step deposition strategy and are highly efficient for multiplex analysis. The cascade amplification composed of RCA and nicking-assisted strand-displacement amplification (SDA) exhibits high amplification efficiency and requires no complicated sequence design, thus improving the compatibility with base-stacking hybridization on our microparticles. Parallel and sensitive analyses for let-7a and miR-21 in one pot without mutual interference have been achieved with both high sensitivity (LOD, approximate to 0.5 fM) and wide dynamic range (10 pM-1 fM). Moreover, our strategy exhibits high specificity for miRNAs of homologous sequence and good anti-interference ability in a complex sample matrix. Considering that there are up to 128 (2(7)) kinds of microparticles available, our strategy can be applied for dozens of miRNA-sensitive analyses in one pot, and it has great potential for miRNA signature analysis as well as widespread clinical applications.