Highly sensitive electrochemical detection of human telomerase activity based on bio-barcode method
Highly sensitive electrochemical detection of human telomerase activity based on bio-barcode method
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DOI：
10.1016/j.bios.2010.04.009
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发表时间：
2010-07-15
影响因子：
12.6
通讯作者：
Wei, Qingli
中科院分区：
文献类型：
--
作者：
Li, Ying;Liu, Bangwei;Wei, Qingli
本研究基于生物条码扩增技术，建立了一种高灵敏度检测人端粒酶活性的电化学方法。从HeLa细胞中提取端粒酶，提取液与端粒酶底物(TS)引物混合进行延伸反应。将延伸产物与固定在金电极上的捕获DNA杂交，然后与金纳米颗粒上的信号DNA反应，形成夹心杂交模式。[Ru(NH(3))(6)](3+)通过静电作用定量结合到Au纳米颗粒上的DNA上，通过计时库仑法产生电化学信号。该方法无需聚合酶链式反应(PCR)扩增端粒酶延伸产物，即可从10个培养的癌细胞中检测到端粒酶活性。(C)2010爱思唯尔B.V.保留所有权利。
In the present study, an electrochemical method for highly sensitive detection of human telomerase activity was developed based on bio-barcode amplification assay. Telomerase was extracted from HeLa cells, then the extract was mixed with telomerase substrate (TS) primer to perform extension reaction. The extension product was hybridized with the capture DNA immobilized on the Au electrode and then reacted with the signal DNA on Au nanoparticles to form a sandwich hybridization mode. Electrochemical signals were generated by chronocoulometric interrogation of [Ru(NH(3))(6)](3+) that quantitatively binds to the DNA on Au nanoparticles via electrostatic interaction. This method can detect the telomerase activity from as little as 10 cultured cancer cells without the polymerase chain reaction (PCR) amplification of telomerase extension product. (C) 2010 Elsevier B.V. All rights reserved.