科研之友
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摘要
Disease marker detection plays an important role in clinical practice; however, rapid, low-cost and simultaneous detection of multiple trace disease markers remain a challenge because of low abundance of related strategies. Herein, we report a G-quadruplex DNAzyme and nicking enzyme-assisted multiplex chemiluminescence (CL) signal amplification (EMCSA) strategy based on a microchip electrophoresis (MCE) platform for simultaneous detection of two trace target molecules. In this study, two target molecules coupled with their hairpin aptamer probes (H-probes), and the open H-probes hybridized with two different hairpin DNAs containing the G-quadruplex sequence (HG4) to form two different DNA duplexes. These DNA duplexes are then selectively cleaved by the nicking enzyme Nb. BbvCI, which forms the target-H-probe complexes and G-rich DNA segments. The obtained target-H-probe complexes hybridize again with another HG4 to form more DNA duplexes, which initiate continuous enzyme cleavage reaction to form more G-rich DNA segments. Free G-rich DNA segments combine with hemin and K+ to form G-quadruplex DNAzymes, which catalyze the CL reaction between hydrogen peroxide (H2O2) and luminol. Finally, G-quadruplex DNAzyme and streptavidin (SA)-functionalised Gquadruplex DNAzyme are quickly separated and detected by the MCE-CL platform. The proposed EMCSA-based MCE-CL method has been applied for simultaneous detection of two significant biomarkers, namely, thrombin (Tb) and carcinoembryonic antigen (CEA), in quantities as low as 2.4 pM and 0.11 ng mL(-1) (0.62 pM), respectively. This method for simultaneous detection of two trace disease markers holds great potential in early clinical diagnosis of diseases.
- 出版日期2017-12-28
- 单位广西师范大学
