Microfluidic circulating reactor system for sensitive and automated duplex-specific nuclease-mediated microRNA detection.
Microfluidic circulating reactor system for sensitive and automated duplex-specific nuclease-mediated microRNA detection.
复制标题
微流控循环反应器系统，用于灵敏和自动的双链特异性核酸酶介导的microRNA检测。
DOI：
10.1016/j.talanta.2021.122396
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发表时间：
2021-09-01
期刊：
影响因子：
6.1
通讯作者：
Zeng, Yong
中科院分区：
文献类型：
--
作者：
Zhou, Xin;Cao, Hongmei;Zeng, Yong
双链特异性核酸酶信号扩增（DSNSA）是一种很有前途的microRNA（miRNA）定量策略。然而，现有的基于DSNSA的miRNA检测方法遭受昂贵的化学品消耗，并且需要费力的多步骤样品预处理，其易于样品损失和污染，包括总RNA提取和富集。为了解决这些问题，本文中，我们设计了一种自动化微流体反应器装置，其将分析物提取/富集和DSNSA介导的miRNA检测集成在一个流线型分析工作流程中。研究了两种流动循环策略，以确定流动条件对珠粒填充微反应器中芯片上DSNSA反应动力学的影响。通过优化的工作流程，我们证明了快速，稳健的芯片上检测miR-21，检测限为35阿莫尔，同时将DSN酶的消耗量大大降低至每次检测0.1 U。因此，这种微流体系统为许多应用提供了有用的工具，包括临床诊断。
Duplex-specific nuclease signal amplification (DSNSA) is a promising microRNA (miRNA) quantification strategy. However, existing DSNSA based miRNA detection methods suffer from costly chemical consumptions and require laborious multi-step sample pretreatment that are prone to sample loss and contamination, including total RNA extraction and enrichment. To address these problems, herein we devised a pneumatically automated microfluidic reactor device that integrates both analyte extraction/enrichment and DSNSA-mediated miRNA detection in one streamlined analysis workflow. Two flow circulation strategies were investigated to determine the effects of flow conditions on the kinetics of on-chip DSNSA reaction in a bead-packed microreactor. With the optimized workflow, we demonstrated rapid, robust on-chip detection of miR-21 with a limit-of-detection of 35 amol, while greatly reducing the consumption of DSN enzyme to 0.1 U per assay. Therefore, this microfluidic system provides a useful tool for many applications, including clinical diagnosis.
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影响因子：
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