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Issue 11, 2013
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Isotachophoresis with ionic spacer and two-stage separation for high sensitivity DNA hybridization assay

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Abstract

We present an on-chip electrophoretic assay for rapid and high sensitivity nucleic acid (NA) detection. The assay uses isotachophoresis (ITP) to enhance NA hybridization and an ionic spacer molecule to subsequently separate reaction products. In the first stage, the probe and target focus and mix rapidly in free solution under ITP. The reaction mixture then enters a region containing a sieving matrix, which allows the spacer ion to overtake and separate the slower probe–target complex from free, unhybridized probes. This results in the formation of two focused ITP peaks corresponding to probe and probe–target complex signals. For a 149 nt DNA target, we achieve a 220 fM limit of detection (LOD) within 10 min, with a 3.5 decade dynamic range. This LOD constitutes a 12× improvement over previous ITP-based hybridization assays. The technique offers an alternative to traditional DNA hybridization assays, and can be multiplexed and extended to detect other biomolecules.

Graphical abstract: Isotachophoresis with ionic spacer and two-stage separation for high sensitivity DNA hybridization assay

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Supplementary files

Article information


Submitted
21 Feb 2013
Accepted
09 Apr 2013
First published
16 Apr 2013

Analyst, 2013,138, 3117-3120
Article type
Communication

Isotachophoresis with ionic spacer and two-stage separation for high sensitivity DNA hybridization assay

C. Eid, G. Garcia-Schwarz and J. G. Santiago, Analyst, 2013, 138, 3117
DOI: 10.1039/C3AN00374D

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