Microflow injection potassium bioassay based on G‐quadruplex DNAzyme‐enhanced chemiluminescence

• DOI: 10.1002/bio.2661
• Publication Date: 2014-05-23T04:32:11Z
• Authors: Lifang Song; Xiaoyan Pan; Hong Shen; Yaling Yu
• Keywords: Reproducibility of Results; DNA, Catalytic; Equipment Design; Hydrogen Peroxide; Aptamers, Nucleotide; Microfluidic Analytical Techniques; 01 natural sciences; 0104 chemical sciences; G-Quadruplexes; Oligodeoxyribonucleotides; Limit of Detection; Flow Injection Analysis; Luminescent Measurements; Potassium; Hemin; Humans; Luminol

ABSTRACTBy taking advantage of microflow injection chemiluminescence analysis, we developed a distinctive microfluidic bioassay method based on G‐Quadruplex DNAzyme‐enhanced chemiluminescence for the determination of K+ in human serum. AGRO100, the G‐rich oligonucleotide with high hemin binding affinity was primarily selected as a K+ recognition element. In the presence of K+, AGRO100 folded into G‐quadruplex and bound hemin to form DNAzyme, which catalyzed the oxidation of luminol by H2O2 to produce chemiluminescence. The intensity of chemiluminescence increased with the K+ concentration. In the study, the DNAzyme showed both long‐term stability and high catalytic activity; other common cations at their physiological concentration did not cause notable interference. With only 6.7 × 10−13 mol of AGRO100 consumption per sample, a linear response of K+ ranged from 1 to 300 µmol/L, the concentration detection limit 0.69 µmol/L (S/N = 3) and the absolute detection limit 1.38 × 10−12 mol were obtained. The precision of 10 replicate measurements of 60 µmol/L K+ was found to be 1.72% (relative standard deviation). The accuracy of the method was demonstrated by analyzing real human serum samples. Copyright © 2014 John Wiley & Sons, Ltd.