Selection and Characterization of a Novel DNA Aptamer for Label-Free Fluorescence Biosensing of Ochratoxin A

Nucleic acid aptamers are emerging as useful molecular recognition tools for food safety monitoring. However, practical and technical challenges limit the number and diversity of available aptamer probes that can be incorporated into novel sensing schemes. This work describes the selection of novel DNA aptamers that bind to the important food contaminant ochratoxin A (OTA). Following 15 rounds of in vitro selection, sequences were analyzed for OTA binding. Two of the isolated aptamers demonstrated high affinity binding and selectivity to this mycotoxin compared to similar food adulterants. These sequences, as well as a truncated aptamer (minimal sequence required for binding), were incorporated into a SYBR® Green I fluorescence-based OTA biosensing scheme. This label-free detection platform is capable of rapid, selective, and sensitive OTA quantification with a limit of detection of 9 nM and linear quantification up to 100 nM

Lateral flow assays for Ochratoxin A using metal nanoparticles: Comparison of "adsorption-desorption" approach to linkage inversion assembled nano-aptasensors (LIANA)

Nano-aptamer probes were prepared and used in lateral flow colorimetric assays for the detection of Ochratoxin A (OTA). In this study, two approaches were examined using 5′-biotin-modified OTA aptamers and silver or gold nanoparticles (AgNP or AuNP). The first method used an "adsorption-desorption" approach wherein aptamers were adsorbed onto the metal nanoparticle surface. Upon the addition of OTA, the aptamer binds specifically to the target, releasing the NPs. The above solutions were applied on a lateral flow assay (LFA) and a detection limit of 6.3 nM was achieved with both metal nanoparticles. The second method used a labelled approach based on Linkage Inversion Assembled Nano-Aptasensors (LIANAs) using a DNA linker containing a 5′-5′ linkage inversion (5′-5′ linker) to assemble biotinylated aptamer-functionalized metal nanoparticles. In the presence of target, OTA specifically binds with its aptamer leading to release of the linker and disassembly of LIANA aggregates into dispersed nanoparticles. The same solutions were applied in LFA format and the lowest detection limit of 0.63 nM was achieved. The results indicated that the LIANA-based LFA strips were more sensitive than the "adsoprtion-desorption" LFAs. Both lateral flow assays are inexpensive, simple, and rapid to perform and produces results visible to the naked-eye

Linkage inversion assembled nano-aptasensors (LIANAs) for turn-on fluorescence detection

A strategy for aptamer-based biosensing termed linkage inversion assembled nano-aptasensors (LIANAs) is shown to be a generally applicable approach to the sensitive and specific detection of a target molecule in turn-on fluorescence solution-based and paper-based tests

Preparation of Multilayered CdSe Quantum Dot Sensitizers by Electrostatic Layer-by-Layer Assembly and a Series of Post-treatments toward Efficient Quantum Dot-Sensitized Mesoporous TiO2 Solar Cells

A multilayer of CdSe quantum dots (QDs) was prepared on the mesoporous surface of a nanoparticulate TiO2 film by a layer-by-layer (LBL) assembly using the electrostatic interaction of the oppositely charged QD surface for application as a sensitizer in QD-sensitized TiO2 solar cells. To maximize the absorption of incident light and the generation of excitons by CdSe QDs within a fixed thickness of TiO2 film, the experimental conditions of QD deposition were optimized by controlling the concentration of salt added into the QD-dissolved solutions and repeating the LBL deposition a few times. A proper concentration of salt was found to be critical in providing a deep penetration of QDs into the mesopore, thus leading to a dense and uniform distribution throughout the whole TiO2 matrix while anchoring the oppositely charged QDs alternately in a controllable way. A series of post-treatments with (1) CdCl2, (2) thermal annealing, and (3) ZnS-coating was found to be very critical in improving the overall photovoltaic properties, presumably through a better connection between QDs, effective passivation of QD's surface, and a high impedance of recombination, which were proved by transmission electron microscopy (TEM) and electrochemical impedance spectroscopy (EIS) experiments. With a proper post-treatment of multilayered QDs as a sensitizer, the overall power conversion efficiency in the CdSe QD-sensitized TiO2 solar cells could reach 1.9% under standard illumination condition of simulated AM 1.5G (100 mW/cm(2)).X113131sciescopu

Fluorescence Enhancement and End-to-End Assembly of Bisacridinedione–Gold Nanorods by Calcium Ions

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Label-free aptasensors based on fluorescent screening assays for the detection of Salmonella typhimurium

We report two label-free fluorescent aptasensor methods for the detection of S. typhimurium. In the first method, we have used a ‘‘turn off’’ approach in which the aptamer is first intercalated with SYBR Green I (SG), leading to a greatly enhanced fluorescence signal. The addition of S. typhimurium (approximately 1530–96938 CFU/mL), which specifically binds with its aptamer and releases SG, leads to a linear decrease in fluorescence intensity. The lowest detection limit achieved with this approach was in the range of 733 CFU/mL. In the second method, a ‘‘turn on’’ approach was designed for S. typhimurium through the Förster resonance energy transfer (FRET) between Rhodamine B (RB) and gold nanoparticles (AuNPs). When the aptamer and AuNPs were mixed with RB, the fluorescence of RB was significantly quenched via FRET. The aptamer adsorbs to the AuNP surface to protect them from salt-induced aggregation, which leads to the fluorescence quenching of RB in presence of AuNPs. Upon the addition of S. typhimurium

Correction to: Comparison of turn-on and ratiometric fluorescent G-quadruplex aptasensor approaches for the detection of ATP (Analytical and Bioanalytical Chemistry, (2019), 10.1007/s00216-018-1484-x)

Regrettably, before online publication the figure of Scheme 2 has been pasted twice as Scheme 1

Comparison of In-Solution Biorecognition Properties of Aptamers against Ochratoxin A

Ochratoxin A (OTA) is a mycotoxin produced as a secondary metabolite by several species of Aspergillus and Penicillium and frequently found as a natural contaminant in a wide range of food commodities. Novel and robust biorecognition agents for detecting this molecule are required. Aptamers are artificial nucleic acid ligands able to bind with high affinity and specificity to a given target molecule. In the last few years, three separate research groups have selected aptamers for ochratoxin A. While each of these three families of aptamers have been incorporated into various methods for detecting OTA, it is unclear if each aptamer candidate is better suited for a particular application. Here, we perform the first head-to-head comparison of solution-based binding parameters for these groups of aptamers. Based on our results, we provide recommendations for the appropriate choice of aptamer for incorporation into solution-based biorecognition assays and applications

Analiza zachowania się rolek z mieszanek kauczukowych napełnionych sadzą w procesie usuwania łuski ryżowej metodą walcowania

In rice husk removal processes, the rubber rollers are used to produce white rice kernels. Paddies are fed between two rubber rollers and the removal of husk takes place due to high friction generated between the paddy layer and outer surface of rubber rollers. Husk removal application involves rubber compounds with promising physico-mechanical properties. Small scale rice mills in Tamil Nadu, India face problems in periodic replacement of rubber rollers due to poor service life. Based on thesurvey, it was found that the rubber compounds with high hardness lead to efficient husk removal and at the same time cause break age of rice. Employing soft rubber eliminates the breakage of rice, but on the other hand the sharp corners of paddy damage the rubber. This research work focuses on the development of rubber compounds for effective rice husk removal application. Carbon black filled, pure and blends of natural rubber (NR), acrylonitrile butadiene rubber (NBR), and epoxidized natural rubber (ENR) are developed. Mechanical properties and lab scale rice husk separation studies are conducted for the developed rubber compounds. The ENR-NBR blends prove to be suitable materials for husk removal application.Usunięcie łusek z ziaren ryżu metodą walcowania pomiędzy rolkami wykonanymi z kauczuku napełnionego sadzą następuje w wyniku tarcia wytwarzanego między warstwą nasion ryżu a powierzchnią rolek. Do tego typu zastosowań potrzebne są mieszanki kauczukowe o specjalnych właściwościach fizyko-mechanicznych. Ze względu na niewielką trwałość dotychczas stosowanych materiałów gumowych, małe młyny ryżu w Tamil Nadu w Indiach borykają się z koniecznością okresowej ich wymiany. Na podstawie przeprowadzonych badań stwierdzono, że rolki wykonane z mieszanek kauczukowych o dużej twardości skutecznie usuwają łuski, ale niestety powodują jednocześnie pękanie ziaren ryżu. Z kolei zastosowanie mieszanek kauczukowych o mniejszej twardości ogranicza zjawisko pękania, ale ostre łuski uszkadzają rolki. Zbadano właściwości mechaniczne i efektywność w procesie separacji łuski ryżowej napełnionych sadzą wulkanizatów kauczuku naturalnego (NR), butadienowo-akrylonitrylowego (NBR), epoksydowanego kauczuku naturalnego (ENR) lub ich mieszanin. Wulkanizaty mieszaniny ENR-NBR okazały się najbardziej odpowiednimi materiałami na rolki do zastosowań w procesie usuwania łuski ryżowej