RESUMEN
Programming ultrasensitive and stimuli-responsive DNAzyme-based probes holds great potential for on-demand biomarker detection. Here, an optically triggered DNAzyme platform was reported for on-demand activation-sensitive electrochemiluminescence (ECL) c-myc mRNA analysis. In this design, the sensing and recognition function of the split DNAzyme (SDz) probe was silent by engineering a blocking sequence containing a photocleavable linker (PC-linker) group at a defined site that could be indirectly cleaved by 302 nm ultraviolet (UV) light. When the SDz probes were assembled on the Au nanoparticles and potassium (K) element doped graphitic carbon nitride nanosheet (K-doped g-C3N4) covered electrode, UV light activation induces the configurational switching and consequently the formation of an active DNAzyme probe with the help of target c-myc mRNA, allowing the cleavage of the substrate strand by magnesium ions (Mg2+). Thus, the release of a ferrocene (Fc)-labeled DNAzyme 2 strand contributed to an extreme ECL signal recovery. In the meantime, the released target c-myc mRNA combined another inactive SDz motif to form active DNAzyme and repeat the cyclic cleavage reaction, resulting in the signal amplification. Furthermore, according to the responses toward two other designed nPC-SDz and m-SDz probes, we demonstrated that controlled UV light mediated photoactivation of the DNAzyme biosensor "on demand" effectively constrained the ECL signal to the mRNA of interest. Moreover, false positive signals could also be avoided due to such a photoactivation design with UV light. Therefore, this study provided a simple methodology that may be broadly applicable for investigating the mRNA-associated physiological events that were difficult to access using traditional DNAzyme probes.
Asunto(s)
ADN Catalítico , Técnicas Electroquímicas , Mediciones Luminiscentes , ARN Mensajero , ADN Catalítico/metabolismo , ADN Catalítico/química , Técnicas Electroquímicas/métodos , ARN Mensajero/análisis , Humanos , Rayos Ultravioleta , Técnicas Biosensibles/métodos , Oro/química , Nanopartículas del Metal/química , Nanopartículas del Metal/efectos de la radiación , Procesos Fotoquímicos , Proteínas Proto-Oncogénicas c-myc/genética , Proteínas Proto-Oncogénicas c-myc/metabolismo , Grafito/química , Límite de Detección , Compuestos de NitrógenoRESUMEN
A green, economical and simple method for the preparation of water-soluble, high-fluorescent carbon quantum dots (CQDs) has been developed via hydrothermal process using pomelo peels as carbon source. The synthesized CQDs were characterized by transmission electron microscopy (TEM), X-ray diffraction(XRD), Fourier transform infrared spectroscopy (FTIR), UV - vis absorption spectra and fluorescence spectrophotometer. The results reveal that the as-prepared C-dots were spherical shape with an average diameter of 2.64 nm and emit bright blue photoluminescence (PL) with a quantum yield of approximately 3.63%. The surface of the C-dots was rich in hydroxyl groups and presented various merits including excellent photostability, low toxicity, and satisfactory solubility. Additionally, we found that two widely used synthetic food colorants, tartrazine and sunset yellow, could result in a strong fluorescence quenching of the C-dots, The possible mechanisms are caused by different ratios of inner filter and static quenching effects. According to this property, This study attempts to establish an analytical method for the determination of tartrazine and sunset yellow using carbon quantum dots as fluorescent probe. A linear relationship was found in the range of 0-100 µM tartrazine and sunset yellow with the detection limit(3σ/k) of 0.65 nM and 1.7 nM. The relative standard deviation (RSD) was 3.5% (tartrazine) and 3.0% (sunset yellow).This observation was further successfully applied for the determination of tartrazine and sunset yellow in food samples collected from local markets, and the recovery rates of the two ranges from 79% to 117.8 and 81 -103.5%, respectively. suggesting its great potential toward food routine analysis.
RESUMEN
A new fluorescent sensor for the determination of lemon yellow was developed based on nitrogen-doped carbon quantum dots (N-CQDs), which were prepared via a hydrothermal method with dried pomelo peel and L-tyrosine. The N-CQDs exhibited the blue fluorescence with a quantum yield of 28 %. The sensing principle of N-CQDs was quenched by lemon yellow via static quenching. The potential interfering substances showed no influence on the detection of lemon yellow. The limit of detection was 0.023 mg/L and lower than that of national standard. Furthermore, the synthesized N-CQDs have been successfully applied to the measurement of lemon yellow in real samples. Hence, the N-CQDs would be a promising sensor in food analysis.
Asunto(s)
Carbono , Nitrógeno , Puntos Cuánticos , Espectrometría de Fluorescencia , Puntos Cuánticos/química , Carbono/química , Nitrógeno/química , Bebidas Gaseosas/análisis , Tecnología Química Verde/métodos , Límite de Detección , Citrus/química , Colorantes Fluorescentes/química , Colorantes Fluorescentes/síntesis químicaRESUMEN
Herein, an electrochemiluminescence (ECL) and electrochemical (EC) dual-mode biosensor platform with a self-powered DNAzyme walking machine was established for accurate and sensitive detection of miRNA-21. By employing a magnesium ion (Mn2+)-dependent DNAzyme cleavage cycling reaction, the walking machine was built by assembling DNAzyme walking strands and ferrocene (Fc)-labeled substrate strands on the Au nanoparticles and graphitic carbon nitride nanosheet (g-C3N4 NS)-covered electrode. The DNAzyme walking strand was first prohibited by a blocker strand. After the addition of target miRNA-21 and Mn2+, the DNAzyme walker could be activated and produce autonomous movements along the electrode track fueled by Mn2+-dependent DNAzyme-catalyzed substrate cleavage without additional energy supply. Notably, each walking step resulted in the cleavage of a substrate strand and the release of a Fc-labeled DNA strand fragment, allowing us to acquire an extreme ECL signal recovery of g-C3N4 inhibited by Fc. Meanwhile, numerous Fc-labeled DNA fragments escaped from the surface of the electrode, directly producing an obvious decrease in the square wave voltammetry (SWV) signal from Fc on the same sensing platform. This work not only avoided difficultly assembling various signal indicators but also significantly improved the sensitivity through using self-powered DNAzyme-walker amplification. Moreover, the proposed design employed the same reaction to produce two signal output modes, which could eliminate the interference from diverse reactive pathways on the outcome to mutually improve the accuracy. Therefore, the dual-mode miRNA-21 biosensor exhibited wide detection ranges of 100 aM to 100 nM with low detection limits of 54.3 and 78.6 aM by ECL and SWV modes, respectively, which provided an efficient and universal biosensing approach with extensive applications in early disease diagnosis and bioanalysis.