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[IEEE 2018 IEEE Life Sciences Conference (LSC) - Montreal, QC, Canada (2018.10.28-2018.10.30)] 2018 IEEE Life Sciences Conference (LSC) - Aptamers-Based Gold-Micro-Array for High-Selective Detection of Bacteria Using Fluorescence Microscopy
摘要： The monitoring of Escherichia coli (E. coli) bacteria in food and water is an effective preventive measure in controlling the outbreaks of serious food-related diseases. Moreover, the accurate detection of its early stage of the bacteria can ensure the public health and safety. In this study, a target-selective aptamer-based platform is proposed as a reliable procedure for the detection of the E. coli in Phosphate-buffered saline (PBS) using fluorescent microscopy. The results show that the aptamer sequences, immobilized on an array of Au-micro-squares, are very high selective towards the target bacteria cells. This platform, presented for the first time, can be potentially used to detect different kinds of cells simultaneously. Also, an important cell-counting application, based on Neubauer Lam high accuracy method, is proposed and implemented and validated by experimental results.
关键词： Fluorescent microscope,Selectivity,E. coli detection,Aptamer
更新于2025-09-23 15:22:29
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Dopamine Binding and Analysis in Undiluted Human Serum and Blood by the RNA-Aptamer Electrode
摘要： Specific analysis of such neurotransmitters as dopamine by the aptamer electrodes in biological fluids is detrimentally affected by non-specific adsorption of media, particularly pronounced at positive charges of the electrode surface at which dopamine oxidizes. Here, we show that dopamine analysis at the RNA-aptamer/cysteamine-modified electrodes is strongly inhibited in undiluted human serum and blood due to non-specific interfacial adsorption of serum and blood components. We demonstrate that non-specific adsorption of serum proteins (but not of blood components) could be minimized when analysis is performed in a flow and injections of serum samples are followed by washing steps in a phosphate buffer solution (PBS) carrier. Under those conditions, the dopamine-aptamer binding affinity in whole human serum of (1.9±0.3)×104 M-1 s-1 was comparable to (3.7±0.3)×104 M-1 s-1 found in PBS, and the dopamine oxidation signal linearly depended on the dopamine concentration, providing the sensitivity of analysis of 73 ± 3 nA μM-1 cm-2 and the LOD of 114 ± 8 nM. The flow-injection apatmer-electrode system was used for direct analysis of basal levels of dopamine in undiluted human serum samples, without using any physical separators (membranes) or filtration procedures. The results suggest a simple strategy for combatting biosurface fouling most pronounced at positive electrode potentials and assist in designing more efficient antifouling strategies for biomedical applications.
关键词： Human serum,Blood,Surface fouling,RNA aptamer electrode,Dopamine,Chronoamperometry,Electrochemical Impedance,Flow-through cell
更新于2025-09-23 15:22:29
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GO-amplified fluorescence polarization assay for high-sensitivity detection of aflatoxin B1 with low dosage aptamer probe
摘要： Aflatoxin B1 (AFB1) is the most toxic mycotoxin of the aflatoxins (AFs) and shows carcinogenic, teratogenic and mutagenic effects in humans and animals. AFB1 is widely seen in cereal products such as rice and wheat. This research proposed a low-cost, high-sensitivity fluorescence polarization (FP) assay for detection of AFB1 using aptamer biosensors based on graphene oxide (GO). The aptamers labelled with fluorescein amidite (FAM) were adsorbed on the surface of GO through π–π stacking and electrostatic interaction, thus forming aptamer/GO macromolecular complexes. Under these conditions, the local rotation of fluorophores was limited and the system had a high FP value. When there was AFB1 in the system, aptamers were dissociated from the GO surface and combined with AFB1 owing to their specificity to form aptamer/AFB1 complexes. As a result, large changes were observed in the molecular weights of aptamers before, and after, the combination, therefore leading to the apparent changes in FP value. The results showed that when only 10 nM of aptamer was used, the changes in FP and the AFB1 concentration had a favourable linear relationship within 0.05 to 5 nM of AFB1, and the lowest detection limit (LOD) was 0.05 nM. In addition, the recoveries of rice sample extract ranged from 89.2% to 112%. The method is simple, highly sensitive, cost-efficient and shows potential application prospects.
关键词： Signal amplifier,Fluorescence polarization,Graphene oxide,Aptamer,Aflatoxin B1
更新于2025-09-23 15:22:29
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[IEEE NAECON 2018 - IEEE National Aerospace and Electronics Conference - Dayton, OH, USA (2018.7.23-2018.7.26)] NAECON 2018 - IEEE National Aerospace and Electronics Conference - Ultrasensitive label-free tobramycin detection with aptamer-functionalized ZnO TFT biosensor
摘要： Aminoglycoside antibiotic such as tobramycin is critical to the treatment of Gram-negative bacterial diseases such as Cystic Fibrosis (Pseudomonas aeruginosa) and other respiratory problems routinely seen in military personnel. Bottom gate (BG) zinc oxide (ZnO) ultra-thin film transistors (TFTs) were fabricated and functionalized with thiol-anchored aptamers as sensitive tobramycin biosensors and a Lower Detection Limit (LDL) of 0.1 nM was extracted from the adsorption coverage isotherm. Sputter deposition of ZnO allows the control of film thickness, stoichiometry, defects and interface states. Intrinsically high surface morphology of ZnO allows improved aptamer functionalization and sensitivity of detection.
关键词： tobramycin,functionalization,biosensor,sputter deposition,aptamer,cystic fibrosis,thin film transistor,ZnO
更新于2025-09-23 15:22:29
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Signal-on Electrochemiluminescence Aptasensor for Bisphenol A based on Hybridization Chain Reaction and Electrically Heated Electrode
摘要： A simple and sensitive electrochemiluminescence (ECL) aptasensor has been developed for bisphenol A (BPA) detection. The capture DNA (CDNA) was modified on the heated indium-tin-oxide (ITO) working electrode surface firstly and then hybridized with BPA aptamer to form double strand DNA (dsDNA). The presence of target can cause the releasing of aptamer from the electrode surface since the aptamer prefers to switch its configuration to combine with BPA. Subsequently, the free CDNA will induce hybridization chain reaction (HCR) to produce long dsDNA on the electrode surface. Ru(phen)3 2+ can integrate into the grooves of dsDNA to act as an ECL reagent, thus enhanced ECL signal can be detected. The temperature control during the processes of target recognition and HCR were realized through the heated electrode instead of the bulk solution heating. Furthermore, the performance of the ECL aptasensor can be further enhanced at elevated electrode temperature. Under the optimized conditions, the ECL intensity of the system has a linear relationship with the logarithm of BPA concentration in the range of 2.0 pM-50 nM. The limit of detection (LOD) at 55 °C (electrode surface temperature) was calculated to be 1.5 pM, which was approximately 6.5-fold lower than that at 25 °C. The proposed biosensor has been applied to detect the BPA in drink samples with satisfactory results.
关键词： electrochemiluminescence,hybridization chain reaction,heated indium-tin-oxide electrode,aptamer,bisphenol A
更新于2025-09-23 15:22:29
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Optical aptasensor based on silver nanoparticles for the colorimetric detection of adenosine
摘要： A new and straightforward optical sensor for the colorimetric determination of adenosine (AD) in human urine samples was developed. The sensor comprised silver nanoparticles (AgNPs) as colorimetric elements and anti-AD aptamer (AP) as a recognition probe. In a solution containing AD and high concentration of NaCl, due to the unique binding of AD with AP, the aggregated metal nanomaterials dispersed in the solution, and the color intensity of the solution was changed accordingly. The absorbance of the solution was monitored for AD quantification. The method was applicable for the determination of AD in the concentration range of 60–280 nM with the detection limit of 21 nM. The relative standard deviation ranged from 4.8 to 8.8% for six replicates. The method showed excellent selectivity toward AD checked over some probable interfering compounds. To investigate the performance of AgNPs, the analytical characteristics of the method including linear range, detection limit, selectivity, and precision were compared with those obtained by a common AuNPs-based aptasensor. The reliability of the method was further ascertained for the detection of AD in urine samples of two lung cancer patients with percentage recoveries in the range of 98–107%.
关键词： Adenosine,Silver nanomaterials,Colorimetric,UV–vis absorption spectroscopy,Aptamer
更新于2025-09-23 15:22:29
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[Environmental Chemistry for a Sustainable World] Nanosensors for Environmental Applications Volume 43 || Development of Optical Sensor Strips for Point-of-Care Testing for Pesticide
摘要： Disposable or point-of-care sensors are a promising tool for low-cost and rapid sensing of analytes including pesticides. They find important applications in pesticide-contaminated food, agro-products, and water quality monitoring. This chapter highlights the implication and significance of pesticide residue identification in foodstuffs and overviews the most frequently engaged analytical techniques, and finally their benefits and limitations are discussed. Disposable strip-based biosensors have their intrinsic advantages and some disadvantages, but their cost-effectiveness and portability have turned them as a potential possibility for point-of-care (POC) testing of various pesticides. The fabrication of robust, low-cost, reliable, and sensitive sensors with the aid of both simple naked eye-based and portable readout-based detectors is the driving factor in this sensor’s technology area. The pending limitations can be overcome by adapting new specific recognition elements and better signal generative particles or systems. The integration of these devices with card readers or smartphones can make them more user-friendly and will provide more accurate quantitative information.
关键词： Organophosphates,Pesticide,Immunoassay,Aptamer,Biosensors,Immunochromatographic assay,Point-of-care,Antibody,Rapid detection,Nanosensors,Gas chromatography
更新于2025-09-23 15:21:01
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Label-free Thrombin Detection Using a Tapered Fiber-optic Interferometric Aptasensor
摘要： Aptamer biosensor for label-free thrombin detection based on a tapered fiber-optic interferometer has been proposed and demonstrated. The biosensor probe can be easily fabricated by tapering a commercial double-cladding fiber down to 5 μm in diameter and it provides a high refractive index sensitivity of 1660 nm/RIU. The changes are determined by tracing the wavelength shift of the dip in the transmission spectrum with variable surrounding refractive index. With the help of surface functionalization of thrombin-binding aptamers, the proposed fiber-optic biosensor proves its capability in label-free detection of thrombin with a lowest detectable concentration of 0.1 μM, together with high specificity in various concentrations of pure and mixed thrombin solutions.
关键词： Fiber-optic biosensor,tapered fiber interferometer,aptamer,thrombin detection
更新于2025-09-23 15:21:01
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Global Folding of a Na+-Specific DNAzyme Studied by FRET
摘要： Recently a few RNA-cleaving DNAzymes have been isolated with excellent specificity for Na+, and some of them contain a Na+ binding aptamer. This metal recognition mechanism is different from most previously reported DNAzymes. Previous studies using 2-aminopurine (2AP) as a probe have indicated interesting local folding by Na+. In this work, fluorescence resonance energy transfer (FRET) was used to probe the global folding of the Ce13d DNAzyme, one of the Na+-specific DNAzymes. FRET pairs were at different locations yielding a total of five constructs to probe its three-way junction structure with a large loop. With end-labelled DNAzymes, the global structure appears quite rigid with little folding by adding up to 200 mM monovalent metal ions, although some minor differences were observed between Li+, Na+ and K+. This lack of large conformational change is also consistent with CD spectroscopy data. The loop was then labelled with an internal TMR fluorophore at the G14 position, and its cleavage activity was partially retained. A clear Na+-dependent folding was observed with spectral crossover. From a biosensing standpoint, global folding based sensors are unlikely to work due to the overall rigid structure of the DNAzyme. Therefore, the best way using this DNAzyme to discriminate Na+ from K+ is based on cleavage activity, followed by probing local folding, while global folding is the least effective for metal discrimination.
关键词： Na+,aptamer,FRET,global folding,DNAzyme
更新于2025-09-23 15:21:01
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A robust covalent coupling scheme for the development of FRET aptasensor based on amino-silane modified graphene oxide
摘要： In recent years, numerous aptamers have been physisorbed on graphene oxide (GO) to develop FRET based aptasensors based on the high fluorescence quenching efficiency of GO. However, physisorbed aptasensors show poor signal reversibility and reproducibility as well as nonspecific probe displacement and thereby, are not suitable for many analytical applications. To overcome these problems when working with complex biological samples, we developed a facile and robust covalent surface functionalization technique for GO-based fluorescent aptasensors using a well studied adenosine triphosphate (ATP) binding aptamer (ABA). In the scheme, GO is first modified with amino-silane, and further with glutaraldehyde to create available carbonyl groups for the covalent attachment of a fluorophore and an amino dual modified ABA. The surface modification method was characterized by zeta potential, X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR). The linearity, sensitivity, selectivity and reversibility of the resulting GO based covalent aptasensor was determined and systematically compared with the physisorbed aptasensor. While both sensors showed similar performance in terms of sensitivity and linearity, better selectivity and higher resistance to nonspecific probe displacement was achieved with the developed covalent ABA sensor. The surface modification technique developed here is independent from the aptamer sequence and therefore could be used universally for different analytical applications simply by changing the aptamer sequence for the target biomolecule.
关键词： aptamer,physisorption,EDC/NHS,fluorescent aptasensor,Graphene oxide,glutaraldehyde,amino-silane,covalent conjugation
更新于2025-09-23 15:21:01