Magnetic particle based electrochemical sensing platform for PCR amplicon detection
Magnetic particle based electrochemical sensing platform for PCR amplicon detection
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DOI：
10.1016/j.elecom.2010.01.036
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发表时间：
2010-04-01
影响因子：
5.4
通讯作者：
Hsing, I-Ming
中科院分区：
文献类型：
--
作者：
Luo, Xiaoteng;Xu, Jingjing;Hsing, I-Ming
本文报道了一种新型的以磁性微粒为分离工具的电化学聚合酶链式反应检测平台。首先将氧化还原活性的插层剂--蒽醌单磺酸(AQMS)插入生物素标记的聚合酶链式反应(PCR)扩增片段中，然后用包被链霉亲和素的磁性颗粒(MPS)捕获形成AQMS-DNA-MP结合物。随后，这些偶联物被吸引到试管的底部，并通过外加磁场从溶液中分离出来，导致溶液AQMS的浓度显著降低。用芯片电极上的差示脉冲伏安法(DPV)监测了溶液中AQMS的浓度变化，以反映PCR扩增片段的存在和数量。考察了模板DNA的初始浓度依赖和循环次数依赖的性能。该电化学聚合酶链式反应检测平台简单、方便、价格低廉，在实际样品监测中具有潜在的应用前景。(C)2010爱思唯尔B.V.保留所有权利。
This work reports a novel electrochemical PCR detection platform using magnetic particles as a separation tool. A redox-active intercalator, anthraquinonemonosulfonic acid (AQMS), was firstly intercalated into biotin labeled PCR amplicons, and the resulting complex was then captured by streptavidin-coated magnetic particles (MPs) to form AQMS-DNA-MP conjugates. Subsequently, these conjugates were attracted to the bottom of the tube and separated from the solution by applying an external magnetic field, resulting in a significant reduction of the concentration of solution AQMS. The concentration changes of solution AQMS, which reflect the presence and quantity of PCR amplicons, were monitored by differential pulse voltammetry (DPV) on a chip electrode. PCR cycle number-dependent as well as the initial template DNA concentration-dependent performances were investigated. This electrochemistry based PCR detection platform is simple, convenient and inexpensive, and may have potential applications for practical sample monitoring. (C) 2010 Elsevier B.V. All rights reserved.