ABSTRACT
In this work, a new electrochemiluminescence (ECL) platform was constructed for detecting the prostate cancer marker microRNA-141 (miRNA-141) on a constant resistor-integrated closed bipolar electrode (BPE). It consisted of two reservoirs and a constant resistor, and both ends were connected to the anode of the driving electrode and the cathode of BPE. The cathode of BPE was modified with boron nitride quantum dots (BNQDs), and the anode reservoir was the [Ru(bpy)3](PF6)2/TPrA system. After introducing a certain amount of hairpin DNA 3 (H3) and ferrocene-labeled single-stranded DNA (Fc-ssDNA) on the surface of the BNQDs, the ECL emission signal of the BNQDs was difficult to be observed by the naked eye, while [Ru(bpy)3](PF6)2 emitted a strong and visible ECL signal. In the presence of the target, bipedal DNA assembled by catalytic hairpin assembly (CHA) took away the Fc-ssDNA and the ECL intensity of the BNQDs was enlarged, and as the concentration of miRNA-141 increased to the cutoff value, yellow-green light was visible by the naked eye. Meanwhile, the red emission signal of [Ru(bpy)3](PF6)2/TPrA became weakened. Thus, an ultrasensitive "color switch" ECL biosensor for detection of miRNA-141 was constructed and endowed with a wide linear range from 10-17 to 10-7 M and a detection limit of 10-17 M (S/N = 3). This study provides the potential for investigating portable devices in the detection of low-concentration nucleic acids.
Subject(s)
Biosensing Techniques , MicroRNAs , DNA , Electrochemical Techniques , Electrodes , Luminescent Measurements , MicroRNAs/analysisABSTRACT
In this article, we developed a novel ECL ratiometry on a closed bipolar electrode (BPE) for the sensitively and accurately detection of miRNA-21. High quantum yield and low toxicity BNQDs was synthesized and coated at BPE cathode as an ECL emitter, while the anode of BPE was calibrated via another ECL material, Ir(df-ppy)2(pic) (Firpic). The electron neutrality at both ends of the BPE electrically coupled the reactions on each pole of the BPE. Therefore, one electrochemical sensing reaction could be quantified at one end of the BPE. By the hybridization of target miRNA-21 and hairpin, the glucose blocked in MSNs by the hairpin was released and reacted with glucose oxidase (GOD) to generate H2O2, thereby reducing the ECL signal of the cathode BNQDs/K2S2O8 system and promoting ECL signal of anode Firpic/TPrA. Further, the G-quadruplex formed by unreacted hairpin bases consumed H2O2, which not only recovered the ECL of BNQDs, but also further improved the ECL emission of Firpic. Therefore, the concentration of miRNA-21 could be measured by the ECL ratio of BNQDs and Firpic. The data showed that the detection limit was 10-15 M (S/N = 3) with the linear range of 10-15 M to 10-9 M. The strategy of the BPE-ECL ratio method based on BNQDs showed a good prospect in clinical application.
