Label-free detection of kanamycin based on a G-quadruplex DNA aptamer-based fluorescent intercalator displacement assay.

This work was the first to report that the kanamycin-binding DNA aptamer (5'-TGG GGG TTG AGG CTA AGC CGA-3') can form stable parallel G-quadruplex DNA (G4-DNA) structures by themselves and that this phenomenon can be verified by nondenaturing polyacrylamide gel electrophoresis and circular dichroism spectroscopy. Based on these findings, we developed a novel label-free strategy for kanamycin detection based on the G4-DNA aptamer-based fluorescent intercalator displacement assay with thiazole orange (TO) as the fluorescence probe. In the proposed strategy, TO became strongly fluorescent upon binding to kanamycin-binding G4-DNA. However, the addition of kanamycin caused the displacement of TO from the G4-DNA-TO conjugate, thereby resulting in decreased fluorescent signal, which was inversely related to the kanamycin concentration. The detection limit of the proposed assay decreased to 59 nM with a linear working range of 0.1 µM to 20 µM for kanamycin. The cross-reactivity against six other antibiotics was negligible compared with the response to kanamycin. A satisfactory recovery of kanamycin in milk samples ranged from 80.1% to 98.0%, confirming the potential of this bioassay in the measurement of kanamycin in various applications. Our results also served as a good reference for developing similar fluorescent G4-DNA-based bioassays in the future.

[Sensing of Cu²âº Based on Fenton Reaction and Unmodified Gold Nanoparticles].

Heavy metal pollution has received great attentions in recent years. The traditional methods for heavy metal detection rely on the expensive laboratory instruments and need time-consuming preparation steps; therefore, it is urgent to develop quick and highly sensitive new technologies for heavy metal detection. The colorimetric method based on the gold nanoparticles (AuNPs) features with simple operation, high sensitivity and low cost, therefore, enabling it widely concerned and used in the environmental monitoring, food safety and chemical and biological sensing fields. This work developed a simple, rapid and highly sensitive strategy based on the Fenton reaction and unmodified AuNPs for the detection of Cu²âº in water samples. The hydroxyl radical ( · OH) generated by the Fenton reaction between the Cu²âº and sodium ascorbate (SA) oxidized the single stranded DNA (ssDNA) attached on the AuNPs surface into variable sequence fragments. The cleavage of ssDNA induced the aggregation of AuNPs in a certain salt solution, therefore, resulting in the changes on the absorbance of solution. The assay conditions were optimized to be pH value of 7.9, 11 mg · L⁻¹ ssDNA, 8 mmol · L⁻¹ SA and 70 mmol · L⁻¹ NaCl. Results showed that the absorbance ratio values at the wavelengths of 700 and 525 nm (A700/A525) were linearly correlated with the Cu²âº concentrations. The linear detection range was 0.1-10.0 µmol · L⁻¹ with a detection limit of 24 nmol · L⁻¹ (3σ). Spiked recoveries ranged from 87%-120% in three sorts of water, including drinking water, tap water and lake water, which confirmed that the potentials of the proposed assay for Cu²âº detection in reality.