Integration of On-Chip Isotachophoresis and Functionalized Hydrogels for Enhanced-Sensitivity Nucleic Acid Detection

被引：53

作者：

Garcia-Schwarz, Giancarlo ^{[1
]}

Santiago, Juan G. ^{[1
]}

机构：

[1] Stanford Univ, Dept Mech Engn, Stanford, CA 94305 USA

来源：

ANALYTICAL CHEMISTRY | 2012年 / 84卷 / 15期

基金：

美国国家科学基金会;

关键词：

EXPRESSION; MICRORNAS; PCR; QUANTIFICATION; CANCERS; RNA; LNA;

D O I：

10.1021/ac301586q

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

We introduce an on-chip electrokinetic assay to perform high-sensitivity nucleic acid (NA) detection. This assay integrates electrokinetic sample focusing using isotachophoresis (ITP) with a background signal-removal strategy that employs photopatterened, DNA-functionalized hydrogels. In this multistage assay, ITP first enhances hybridization kinetics between target NAs and end-labeled complementary reporters. After enhanced hybridization, migration through a DNA-functionalized hydrogel region removes excess reporters through affinity interactions. We demonstrate our assay on microRNAs, an important class of low-abundance biomarkers. The assay exhibits 4 orders of magnitude dynamic range, near 1 pM detection limits starting from less than 100 fg of microRNA, and high selectivity for mature microRNA sequences, all within a 10 min run time. This new microfluiclic framework provides a unique quantitative assay for NA detection.

引用

页码：6366 / 6369

页数：4

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