Electrochemical detection of low-copy number salivary RNA based on specific signal amplification with a hairpin probe.
Electrochemical detection of low-copy number salivary RNA based on specific signal amplification with a hairpin probe.
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DOI：
10.1093/nar/gkn299
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发表时间：
2008-06
影响因子：
14.9
通讯作者：
Ho CM
中科院分区：
文献类型：
--
作者：
Wei F;Wang J;Liao W;Zimmermann BG;Wong DT;Ho CM
我们开发了一种利用发夹探针（HP）电化学检测唾液mRNA的技术。空间位阻（SH）抑制非特异性信号并产生信号放大过程，用于目标检测。茎环结构使探针的报告端靠近表面，使其无法与中介体结合。目标绑定打开探针的发夹结构，然后中介可以绑定到可访问的报告器。利用辣根过氧化物酶产生电化学信号。这种信号接通过程的特点是低基础信号，强正读数和大动态范围。SH通过发夹设计和电场控制。通过对HPs施加电场控制，RNA的检测限约为0.4 fM，比传统的线性探针灵敏度高10000倍。HP检测内源性白细胞介素-8 mRNA，与定量PCR技术有较好的相关性。由此产生的过程允许一个简单的设置，并通过减少步骤的数量，它适用于点护理检测特定的核酸序列从复杂的体液，如唾液。
We developed a technique for electrochemical detection of salivary mRNA employing a hairpin probe (HP). Steric hindrance (SH) suppresses unspecific signal and generates a signal-on amplification process for target detection. The stem-loop configuration brings the reporter end of the probe into close proximity with the surface and makes it unavailable for binding with the mediator. Target binding opens the hairpin structure of the probe, and the mediator can then bind to the accessible reporter. Horseradish peroxidase is utilized to generate electrochemical signal. This signal-on process is characterized by a low basal signal, a strong positive readout and a large dynamic range. The SH is controlled via hairpin design and electrical field. By applying electric field control to HPs, the limit of detection of RNA is about 0.4 fM, which is 10 000-fold more sensitive than conventional linear probes. Endogenous Interleukin-8 mRNA is detected with the HP, and good correlation with the quantitative PCR technique is obtained. The resultant process allows a simple setup and by reducing the number of steps it is suited for the point-of-care detection of specific nucleic acid sequences from complex body fluids such as saliva.
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影响因子：
9.4
作者：
Liao, JC;Mastali, M;Haake, DA
通讯作者：
Haake, DA
影响因子：
3.4
作者：
Rant, U;Arinaga, K;Abstreiter, G
通讯作者：
Abstreiter, G
影响因子：
3.9
作者：
Su, XD;Wu, YJ;Knoll, W
通讯作者：
Knoll, W
影响因子：
12.6
作者：
Wei, F;Sun, B;Zhao, XS
通讯作者：
Zhao, XS
DOI：
10.14219/jada.archive.1993.0007
发表时间：
1993-01-01
影响因子：
3.9
作者：
MANDEL, ID
通讯作者：
MANDEL, ID