Abstract

Coupling isotachophoresis with bead-based assay for rapid and multiplexed nucleic acids detection

Hirofumi Shintaku, Assistant Professor, Kyoto University

We present an electrokinetic technique to increase the reaction rate and sensitivity of multiplexed bead-based nucleic acid assay. We use isotachophoresis (ITP) to co-focus and preconcentrate target molecules and beads functionalized with conjugate probe molecules into a single ITP zone. We here demonstrate quantitative detection of 24 nt single-stranded DNA over a dynamic range of three orders of magnitude and multiplexed detection of ten target species per sample. We show that ITP can achieve approximately the same sensitivity in 60-fold less reaction time (20 min versus 20 h). Alternately, compared to standard minimum reaction time of 30 min, we show that 20 min ITP hybridization can achieve 5.3-fold higher sensitivity. (H. Shintaku et al. Angew Chem Int Ed, 2014)