In this work, Ag nanoparticles (AgNPs) were synthesized quickly by a one-step method utilizing polydopamine-glutathione nanoparticles (PDA-GNPs) as a reducing agent. The PDA-GNPs and the generated AgNPs acted as the energy donor and acceptor, respectively. Accordingly, the fluorescence of PDA-GNPs was quenched on the basis of fluorescence resonance energy transfer (FRET). In the presence of melamine, the preferential combination of Ag(I) and melamine to form Ag(I)-melamine complex prevents Ag(I) from forming AgNPs, together with fluorescence enhancement compared with the absence of melamine. Under the optimal conditions including the concentration of AgNO, reaction time, reaction temperature, and pH, the fluorescence enhancement efficiency has a linear response to the concentration of melamine from 0.1 to 40 μM with a detection limit of 23 nM for melamine. The proposed method is simple, time-saving, and low-cost, which was further applied to detect melamine in real milk products with satisfactory results.
ExperimentalReagents 3,6-Dimethyl-2-(4-dimethylaminophenyl) benzo-thiazolium cation (ThT), potassium chloride (KCl), and magnesium chloride (MgCl2) were purchased from Sangon Inc. (Shanghai, China).2-Amino-2-(hydroxymethyl)-1,3-propanediol (Tris) The detection of disease-related DNA is of great significance for early and accurate diagnosis and therapy. In this work, we successfully achieved the sensitive detection of target DNA based on a thioflavin T (ThT)-induced G-quadruplex fluorescent biosensor. ThT, a water-soluble fluorescent dye, can induce G-rich sequences to form G-quadruplexes and obtain an obviously enhanced fluorescence. In this work, it was employed to construct a biosensor for the detection of HIV. When the target HIV existed, the hairpin DNA probes would be opened in succession and release the completely exposed G-rich sequence to combine with ThT. The simple and rapid biosensor performed satisfactory selectivity; it also exhibited sensitivity with a detection limit of 2.4 nM. With good performance in human serum, we believe that this optical biosensor has the potential to be applied to the practical detection of target DNA.
The use of alkaline phosphatase (ALP) as a biomarker in some diseases including hepatitis, obstructive jaundice, osteoblastic bone cancer, and osteomalacia is important in clinical diagnosis. Furthermore, ALP activity detection is an essential hot topic in environmental monitoring, biomedical research, and other research fields. In this study, a novel "signal-on" photoelectrochemical (PEC) biosensor based on the ALP-catalyzed phosphorylation reaction was designed to sensitively detect ALP activity. In this design, ascorbic acid-an electron donor-was catalytically produced by ALP from l-ascorbic acid 2-phosphate trisodium salt in situ, which results in an increased photocurrent response signal. For immobilizing the ALP on the electrode surface, poly diallyl dimethyl ammonium chloride was used for the conjugation of ALP, and titanium dioxide (TiO)-a photoactive material-and graphite-like carbon nitride (g-CN) nanocomposites were prepared and characterized. TiO attached on g-CN plays an important role for the biosensing purpose due to their good biocompatibility and chemical/thermal stability, while g-CN provides the PEC response signal. Furthermore, the prepared TiO/g-CN nanocomposites can effectively suppress electron-hole recombinations, improve the excitation conversion efficiency, and make the best use of solar energy. The PEC biosensor for ALP activity detection displays a detection limit of 0.03 U L (S/N = 3), which offers a new route for the ALP activity assay in human serum samples.
Based on the toehold-mediated strand displacement reaction and exonuclease III assisted amplification, a sensitive and simple target DNA biosensor was established.
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