4.8 Article

Detection of microRNA by Electrocatalytic Amplification: A General Approach for Single-Particle Biosensing

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JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
卷 139, 期 22, 页码 7657-7664

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AMER CHEMICAL SOC
DOI: 10.1021/jacs.7b03648

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  1. U.S. Defense Threat Reduction Agency [HDTRA1-11-1-0005]
  2. Robert A. Welch Foundation [F-0032]

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Here we report a sensing scheme for detection of microRNA (miRNA) using electrocatalytic amplification (ECA). ECA is a method in which nanoparticles (NPs) that are catalytic for a specific electrochemical reaction collide with an inert electrode surface. Each collision results in a detectable current transient. In the present article, we show that this general approach can be extended to detection of miRNA. Specifically, PtNPs are modified with a single-strand DNA (ssDNA) shell that is complementary to the miRNA target. Next, the ssDNA:miRNA conjugate is formed, which passivates the PtNP surface. In the presence of an enzyme called duplex specific nuclease (DSN), however, a fraction of the surface-bound DNA is removed thereby exposing some of the PtNP surface. In other words, the electrocatalytic properties of the PtNPs are reactivated only if miRNA complementary to ssDNA is present. This methodology resolves a number of problems that have rendered ECA ineffective for biosensing applications. Moreover, the results suggest that the underlying chemistry is broadly applicable to nucleic acid sensing.

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