Nanopore-based aptasensor for label-free and sensitive vanillin determination in food samples.

Dielectric breakdown technique was utlised to fabricate 5-6 nm nanopores for vanillin detection in various food samples. A highly selective aptamer (Van_74) with high binding affinity towards vanillin was used as capture probe. Under optimal conditions, aptamer/vanillin complex translocation induced deeper events than the bare aptamer. As a result, the proposed nanopore aptasensor exhibits a linear range from 0.5 to 5 nM (R2 = 0.972) and a low detection limit of 500 pM, which is significantly better than conventional platforms. Furthermore, our aptasensor showed excellent immunity against different interferons and was used to detect vanillin in different food samples. The food sample measurements were confirmed with an additional UV-Vis assay, the results of the two techniques were statistically evaluated and showed no statistically significant difference. Hence, this work represents a proof-of-concept involving the design and testing of aptamer/nanopore sensors for small molecules detection, which plays a critical role in food safety.

Highly sensitive fluorescence multiplexed miRNAs biosensors for accurate clinically diagnosis lung cancer disease using LNA-modified DNA probe and DSN enzyme.

With the emergence of microRNAs as key biomarkers for disease diagnosis such as lung cancer, various techniques have been settled for their detection. However, these current methods require different amplification steps since numerous challenges for detecting circulating miRNAs are attributable to their intrinsic properties accounting for tiny sizes, high sequence similarity, and low abundance. Duplex specific nuclease (DSN)-based microRNA amplification has recently gained interest in biosensing applications thanks to its catalytic activity based on target recycling. In this context, we designed a highly selective, sensitive, and multiplexed fluorescence-based biosensor combining DSN enzyme and magnetic beads to detect three distinct microRNAs, including microRNA-21, microRNA-210, and microRNA-486-5p. By exploiting the above approach, we were able to detect as low as 98 aM, 120 aM, and 300 aM of mir-21, miR-210, and miR-486-5p, respectively. Furthermore, this recommended strategy displays a high selectivity toward an entirely matched target than the off-target. These results are ascribed to the potent DSN enzyme activity and to the locked nucleic acid (LNA)-modified DNA probe that boosted the hetero-duplex probe/target stability. Lastly, our proposed method was applied to detect microRNAs in the serum samples and displayed a high efficacy to discriminate between healthy controls and lung cancer patients. Furthermore, the analytical accuracy of the proposed strategy was validated with the computed tomography (CT) technique of the chest. Thus based on these findings, this strategy could open new directions for detecting microRNAs associated with several diseases.

Highly Sensitive Fluorescence Assay for miRNA Detection: Investigation of the DNA Spacer Effect on the DSN Enzyme Activity toward Magnetic-Bead-Tethered Probes.

Researchers have recently designed various biosensors combining magnetic beads (MBs) and duplex-specific nuclease (DSN) enzyme to detect miRNAs. Yet, the interfacial mechanisms for surface-based hybridization and DSN-assisted target recycling are relatively not well understood. Thus, herein, we developed a highly sensitive and selective fluorescent biosensor to study the phenomenon that occurs on the local microenvironment surrounding the MB-tethered DNA probe via detecting microRNA-21 as a model. Using the above strategy, we investigated the influence of different DNA spacers, base-pair orientations, and surface densities on DSN-assisted target recycling. As a result, we were able to detect as low as 170 aM of miR-21 under the optimized conditions. Moreover, this approach exhibits a high selectivity in a fully matched target compared to a single-base mismatch, allowing the detection of miRNAs in serum with improved recovery. These results are attributed to the synergetic effect between the DSN enzyme activity and the neutral DNA spacer (triethylene glycol: TEG) to improve the miRNA detection's sensitivity. Finally, our strategy could create new paths for detecting microRNAs since it obliterates the enzyme-mediated cascade reaction used in previous studies, which is more expensive, more time-consuming, less sensitive, and requires double catalytic reactions.

Facile and Controllable Synthesis of Large-Area Monolayer WS2 Flakes Based on WO3 Precursor Drop-Casted Substrates by Chemical Vapor Deposition.

Monolayer WS2 (Tungsten Disulfide) with a direct-energy gap and excellent photoluminescence quantum yield at room temperature shows potential applications in optoelectronics. However, controllable synthesis of large-area monolayer WS2 is still challenging because of the difficulty in controlling the interrelated growth parameters. Herein, we report a facile and controllable method for synthesis of large-area monolayer WS2 flakes by direct sulfurization of powdered WO3 (Tungsten Trioxide) drop-casted on SiO2/Si substrates in a one-end sealed quartz tube. The samples were thoroughly characterized by an optical microscope, atomic force microscope, transmission electron microscope, fluorescence microscope, photoluminescence spectrometer, and Raman spectrometer. The obtained results indicate that large triangular monolayer WS2 flakes with an edge length up to 250 to 370 µm and homogeneous crystallinity were readily synthesized within 5 min of growth. We demonstrate that the as-grown monolayer WS2 flakes show distinctly size-dependent fluorescence emission, which is mainly attributed to the heterogeneous release of intrinsic tensile strain after growth.

Label-free identification of trace microcystin-LR with surface-enhanced Raman scattering spectra.

The analysis of trace microcystin-LR (MC-LR) plays important roles in environmental fields, especially in monitoring domestic water quality and safety, since it has particularly harmful effect on wild and domestic animals as well as humans at low doses. Herein, we combine confocal Raman spectroscopy with SERS-AG substrate to characterize the "fingerprint" information of MC-LR directly. High sensitivity of SERS-AG substrates was verified by utilizing the probe molecule Rhodamine 6 G. Mapping spectra demonstrated good reproducibility of MC-LR identification with label-free surface-enhanced Raman scattering (SERS) strategy. Differences between SERS spectra of MC-LR and R6G, microcystin-RR were evaluated by calculating their scores and loading weights with an unsupervised exploratory principal component analysis method. Then, relationship between Raman intensities and concentrations was preliminary analyzed with SERS spectra of MC-LR and the lowest concentration of MC-LR identification was 10-6 mg L-1 while using SERS-AG substrate. Thereafter, 68.6% quantitative recovery of 10-3 mg L-1 MC-LR in tap water samples was obtained by the proposed label-free SERS method. These results showed that confocal Raman spectroscopy with label-free surface-enhanced Raman scattering strategy can handle the identification of trace MC-LR for monitoring water quality and safety worldwide in future.