scholarly journals A Ratiometric Fluorescent Aptamer Homogeneous Biosensor Based on Hairpin Structure Aptamer for AFB1 Detection

2022 ◽  
Author(s):  
Beibei Feng ◽  
Fei Zhao ◽  
Min Wei ◽  
Yong Liu ◽  
Xinyu Ren ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Resonance Energy Transfer ◽  
Fluorescence Emission ◽  
Resonance Energy ◽  
Hairpin Structure ◽  
Fluorescence Signal ◽  
Detection Range ◽  
Quenching Effect ◽  
Double Stranded Dna ◽  
Aflatoxin B

Abstract On the basis of aptamer (Apt) with hairpin structure and fluorescence resonance energy transfer (FRET), a ratio fluorescent aptamer homogeneous sensor was prepared for the determination of Aflatoxin B1 (AFB1). Initially, the Apt labeled simultaneously with Cy5, BHQ2, and cDNA labeled with Cy3 were formed a double-stranded DNA through complementary base pairing. The fluorescent aptamer sensor demonstrates a weak fluorescence emission of Cy3 and a high fluorescence emission of Cy5 due to the quenching effect of BHQ2. The double-stranded DNA structure will be disintegrated in the presence of AFB1, resulting the removal of Cy3 and the close of Cy5 with BHQ2. The fluorescence signal of Cy3 and Cy5 were restored and quenched respectively. Thus, the ratio change of FCy3 to FCy5 was used to realized the detection of AFB1 with wider detection range and lower limit of detection (LOD). The response of the optimized protocol for AFB1 detection was wider linear range from 0.05 ng/mL to 100 ng/mL and the LOD was 12.6 pg/mL. The sensor designed in this strategy has the advantages of simple preparation and fast signal response. It has been used for the detection of AFB1 in labeled corn and wine, indicating it had good application potential in practical samples.

scholarly journals A FRET-ICT Dual-Modulated Ratiometric Fluorescence Sensor for Monitoring and Bio-Imaging of Cellular Selenocysteine

Molecules ◽  
2020 ◽  
Vol 25 (21) ◽  
pp. 4999
Author(s):  
Zongcheng Wang ◽  
Chenhong Hao ◽  
Xiaofang Luo ◽  
Qiyao Wu ◽  
Chengliang Zhang ◽  
...  
Keyword(s):  
Real Time ◽  
Limit Of Detection ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
High Sensitivity ◽  
Fluorescence Sensor ◽  
Quantitative Detection ◽  
Fluorescence Signal ◽  
Ratiometric Fluorescence ◽  
Sensitivity And Selectivity

Since the fluctuation of cellular selenocysteine (Sec) concentration plays an all-important role in the development of numerous human disorders, the real-time fluorescence detection of Sec in living systems has attracted plenty of interest during the past decade. In order to obtain a faster and more sensitive small organic molecule fluorescence sensor for the Sec detection, a new ratiometric fluorescence sensor Q7 was designed based on the fluorescence resonance energy transfer (FRET) strategy with coumarin fluorophore as energy donor and 4-hydroxy naphthalimide fluorophore (with 2,4-dinitrobenzene sulfonate as fluorescence signal quencher and Sec-selective recognition site) as an energy acceptor. The sensor Q7 exhibited only a blue fluorescence signal, and displayed two well distinguished emission bands (blue and green) in the presence of Sec with ∆λ of 68 nm. Moreover, concentrations ranging of quantitative detection of Sec of Q7 was from 0 to 45 μM (limit of detection = 6.9 nM), with rapid ratiometric response, high sensitivity and selectivity capability. Impressively, the results of the living cell imaging test demonstrated Q7 has the potentiality of being an ideal sensor for real-time Sec detection in biosystems.

scholarly journals An Aptamer-Array-Based Sample-to-Answer Biosensor for Ochratoxin A Detection via Fluorescence Resonance Energy Transfer

Chemosensors ◽  
2021 ◽  
Vol 9 (11) ◽  
pp. 309
Author(s):  
Yongning Li ◽  
Zhenfei Peng ◽  
Yaxi Li ◽  
Min Xiao ◽  
Gongjun Tan ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Ochratoxin A ◽  
Fluorescence Resonance Energy Transfer ◽  
Limit Of Detection ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
High Sensitivity ◽  
Detection Methods ◽  
Fluorescence Signal ◽  
Fluorescence Resonance

Food toxins are a hidden threat that can cause cancer and tremendously impact human health. Therefore, the detection of food toxins in a timely manner with high sensitivity is of paramount importance for public health and food safety. However, the current detection methods are relatively time-consuming and not practical for field tests. In the present work, we developed a novel aptamer-chip-based sample-to-answer biosensor (ACSB) for ochratoxin A (OTA) detection via fluorescence resonance energy transfer (FRET). In this system, a cyanine 3 (Cy3)-labeled OTA-specific biotinylated aptamer was immobilized on an epoxy-coated chip via streptavidin-biotin binding. A complementary DNA strand to OTA aptamer at the 3′-end was labeled with a black hole quencher 2 (BHQ2) to quench Cy3 fluorescence when in proximity. In the presence of OTA, the Cy3-labeled OTA aptamer bound specifically to OTA and led to the physical separation of Cy3 and BHQ2, which resulted in an increase of fluorescence signal. The limit of detection (LOD) of this ACSB for OTA was 0.005 ng/mL with a linearity range of 0.01–10 ng/mL. The cross-reactivity of ACSB against other mycotoxins, ochratoxin B (OTB), aflatoxin B1 (AFB1), zearalenone (ZEA), or deoxynilvalenol (DON), was less than 0.01%. In addition, this system could accurately detect OTA in rice samples spiked with OTA, and the mean recovery rate of the spiked-in OTA reached 91%, with a coefficient of variation (CV) of 8.57–9.89%. Collectively, the ACSB may represent a rapid, accurate, and easy-to-use platform for OTA detection with high sensitivity and specificity.

scholarly journals A Compact Detection Platform Based on Gradient Guided-Mode Resonance for Colorimetric and Fluorescence Liquid Assay Detection

Sensors ◽  
2021 ◽  
Vol 21 (8) ◽  
pp. 2797
Author(s):  
Jing-Jhong Gao ◽  
Ching-Wei Chiu ◽  
Kuo-Hsing Wen ◽  
Cheng-Sheng Huang
Keyword(s):  
Limit Of Detection ◽  
Detection System ◽  
Fluorescence Emission ◽  
Assay Sensitivity ◽  
Detection Range ◽  
Current Form ◽  
Guided Mode ◽  
Spectral Detection ◽  
Guided Mode Resonance ◽  
Colorimetric Assays

This paper presents a compact spectral detection system for common fluorescent and colorimetric assays. This system includes a gradient grating period guided-mode resonance (GGP-GMR) filter and charge-coupled device. In its current form, the GGP-GMR filter, which has a size of less than 2.5 mm, can achieve a spectral detection range of 500–700 nm. Through the direct measurement of the fluorescence emission, the proposed system was demonstrated to detect both the peak wavelength and its corresponding intensity. One fluorescent assay (albumin) and two colorimetric assays (albumin and creatinine) were performed to demonstrate the practical application of the proposed system for quantifying common liquid assays. The results of our system exhibited suitable agreement with those of a commercial spectrometer in terms of the assay sensitivity and limit of detection (LOD). With the proposed system, the fluorescent albumin, colorimetric albumin, and colorimetric creatinine assays achieved LODs of 40.99 and 398 and 25.49 mg/L, respectively. For a wide selection of biomolecules in point-of-care applications, the spectral detection range achieved by the GGP-GMR filter can be further extended and the simple and compact optical path configuration can be integrated with a lab-on-a-chip system.

scholarly journals Ratiometric Fluorescent Biosensors for Glucose and Lactate Using an Oxygen-Sensing Membrane

Biosensors ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 208
Author(s):  
Hong Dinh Duong ◽  
Jong Il Rhee
Keyword(s):  
Limit Of Detection ◽  
Oxygen Sensing ◽  
High Sensitivity ◽  
Glucose Sensing ◽  
Oxidation Reactions ◽  
Lactate Oxidase ◽  
Lactate Oxidation ◽  
Detection Range ◽  
Quenching Effect ◽  
Sensing Membrane

In this study, ratiometric fluorescent glucose and lactate biosensors were developed using a ratiometric fluorescent oxygen-sensing membrane immobilized with glucose oxidase (GOD) or lactate oxidase (LOX). Herein, the ratiometric fluorescent oxygen-sensing membrane was fabricated with the ratio of two emission wavelengths of platinum meso-tetra (pentafluorophenyl) porphyrin (PtP) doped in polystyrene particles and coumarin 6 (C6) captured into silica particles. The operation mechanism of the sensing membranes was based on (i) the fluorescence quenching effect of the PtP dye by oxygen molecules, and (ii) the consumption of oxygen levels in the glucose or lactate oxidation reactions under the catalysis of GOD or LOX. The ratiometric fluorescent glucose-sensing membrane showed high sensitivity to glucose in the range of 0.1–2 mM, with a limit of detection (LOD) of 0.031 mM, whereas the ratiometric fluorescent lactate-sensing membrane showed the linear detection range of 0.1–0.8 mM, with an LOD of 0.06 mM. These sensing membranes also showed good selectivity, fast reversibility, and stability over long-term use. They were applied to detect glucose and lactate in artificial human serum, and they provided reliable measurement results.

scholarly journals Graphene Oxide and Fluorescent Aptamer Based Novel Biosensor for Detection of 25-hydroxyvitamin D3

2021 ◽  
Author(s):  
Ritika Gupta ◽  
Sunaina Kaul ◽  
Vishal Singh ◽  
Sandeep Kumar ◽  
Nitin Kumar Singhal
Keyword(s):  
Vitamin D ◽  
Graphene Oxide ◽  
Limit Of Detection ◽  
Low Cost ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Detection Methods ◽  
Hydroxyvitamin D ◽  
25 Hydroxyvitamin D ◽  
Sensitive Sensor

Abstract For maintaining the healthy metabolic status, vitamin D is a beneficial metabolite stored majorly in its pre-activated form, 25-hydroxyvitamin D3 (25(OH)D3). Due to its important role in bone strengthening, the study was planned to quantify 25(OH)D3 levels in our blood. Quantification techniques for 25(OH)D3 are costly thus requiring a need for a low cost, and sensitive detection methods. In this work, an economic, and sensitive sensor for the detection of 25(OH)D3 was developed using aptamer and graphene oxide (GO). Aptamer is an oligonucleotide, sensitive towards its target, whereas, GO with 2D nanosheets provides excellent quenching surface. Aptamer labeled with fluorescein (5’, 6-FAM) is adsorbed by π -π interaction on the GO sheets leading to quenching of the fluorescence due to Förster resonance energy transfer (FRET). However, in the presence of 25(OH)D3, a major portion of aptamer fluorescence remains unaltered, due to its association with 25(OH)D3. However, in the absence, aptamer fluorescence gets fully quenched. Fluorescence intensity quenching was monitored using fluorescence spectrophotometer and agarose gel based system. The limit of detection of 25(OH)D3 by this method was found to be 0.15 µg/mL. Therefore, this method could come up as a new sensing method in the field of vitamin D detection.

scholarly journals Affinity series of genetically encoded high sensitivity Förster Resonance Energy Transfer sensors for sucrose

2020 ◽  
Author(s):  
Mayuri Sadoine ◽  
Mira Reger ◽  
Ka Man Wong ◽  
Wolf B. Frommer
Keyword(s):  
Energy Transfer ◽  
Steady State ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Forster Resonance Energy Transfer ◽  
Förster Resonance Energy Transfer ◽  
Detection Range ◽  
Steady State Levels ◽  
Förster Resonance

ABSTRACTGenetically encoded fluorescent sugar sensors are valuable tools for the discovery of transporters and for quantitative monitoring of sugar steady-state levels in intact tissues. Genetically encoded Förster Resonance Energy Transfer sensors for glucose have been designed and optimized extensively, and a full series of affinity mutants is available for in vivo studies. However, to date, only a single improved sensor FLIPsuc-90µΔ1 with a Km for sucrose of ∼90 µM is available for sucrose monitoring. This sucrose sensor was engineered on the basis of an Agrobacterium tumefaciens sugar binding protein. Here, we took a two-step approach to first systematically improve the dynamic range of the FLIPsuc nanosensor and then expand the detection range from micromolar to millimolar sucrose concentrations by mutating a key residue in the binding site. The resulting series of sucrose sensors may allow systematic investigation of sucrose transporter candidates and comprehensive in vivo analyses of sucrose concentration in plants. Since FLIPsuc-90µ also detects trehalose in animal cells, the new series of sensors can be used to investigate trehalose transporter candidates and monitor trehalose steady-state levels in vivo as well.

scholarly journals Peptide-coated palladium nanoparticle for highly sensitive bioanalysis of trypsin in human urine samples

2018 ◽  
Vol 8 ◽  
pp. 184798041882039 ◽  
Author(s):  
Guohua Zhou ◽  
Huimin Jiang ◽  
Yanfang Zhou ◽  
Peilian Liu ◽  
Yongmei Jia ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Palladium Nanoparticles ◽  
Limit Of Detection ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Water Soluble ◽  
Urine Samples ◽  
Fluorescence Resonance ◽  
Functional Peptide

In recent years, palladium nanoparticles have been proved as energy acceptor candidates in fluorescence resonance energy transfer-based sensors for analytical and biological purposes. In this article, peptide-coated palladium nanoparticles were prepared using a simple one-step preparation method. The peptide Cys-Ala-Leu-Asn-Asn was used as a ligand, whereas hydrazine hydrate was used as a reductant to obtain water-soluble and stable peptide-coated palladium nanoparticles. Additionally, peptide-coated palladium nanoparticles were functionalized by adding the functional peptide CALNNGGARK(FITC) in combination with Cys-Ala-Leu-Asn-Asn during the preparation process. The prepared functionalized peptide-coated palladium nanoparticles were used for trypsin detection based on the fluorescence resonance energy transfer approach. Under optimized conditions, the proposed method can be used for the detection of trypsin concentrations in the range of approximately 0.2–8-μg/mL with a limit of detection of 0.18-μg/mL. The functionalized peptide-coated palladium nanoparticles were successfully applied for the detection of trypsin in urine samples. Our findings also indicated that peptide-coated palladium nanoparticles can highly quench fluorophores and are suitable for the manufacture of off–on state fluorescent sensors. We anticipated that the peptide-coated palladium nanoparticles proposed in this article will have great potential for the detection of trypsin in urine and other analytical, biological, and clinical applications.

scholarly journals A novel fluorescent biosensor based on dendritic DNA nanostructure in combination with ligase reaction for ultrasensitive detection of DNA methylation

2019 ◽  
Vol 17 (1) ◽  
Author(s):  
Shu Zhang ◽  
Jian Huang ◽  
Jingrun Lu ◽  
Min Liu ◽  
Yan Li ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Limit Of Detection ◽  
Fluorescent Dye ◽  
Fluorescence Signal ◽  
Target Sequence ◽  
Capture Probe ◽  
Detection Range ◽  
Fluorescent Biosensor ◽  
Dna Nanostructure ◽  
Reporter Probe

Background DNA methylation detection is indispensable for the diagnosis and prognosis of various diseases including malignancies. Hence, it is crucial to develop a simple, sensitive, and specific detection strategy. Methods A novel fluorescent biosensor was developed based on a simple dual signal amplification strategy using functional dendritic DNA nanostructure and signal-enriching polystyrene microbeads in combination with ligase detection reaction (LDR). Dendritic DNA self-assembled from Y-DNA and X-DNA through enzyme-free DNA catalysis of a hairpin structure, which was prevented from unwinding at high temperature by adding psoralen. Then dendritic DNA polymer labeled with fluorescent dye Cy5 was ligated with reporter probe into a conjugate. Avidin-labeled polystyrene microbeads were specifically bound to biotin-labeled capture probe, and hybridized with target sequence and dendritic DNA. LDR was triggered by adding Taq ligase. When methylated cytosine existed, the capture probe and reporter probe labeled with fluorescent dye perfectly matched the target sequence, forming a stable duplex to generate a fluorescence signal. However, after bisulfite treatment, unmethylated cytosine was converted into uracil, resulting in a single base mismatch. No fluorescence signal was detected due to the absence of duplex. Results The obtained dendritic DNA polymer had a large volume. This method was time-saving and low-cost. Under the optimal experimental conditions using avidin-labeled polystyrene microbeads, the fluorescence signal was amplified more obviously, and DNA methylation was quantified ultrasensitively and selectively. The detection range of this sensor was 10−15 to 10−7 M, and the limit of detection reached as low as 0.4 fM. The constructed biosensor was also successfully used to analyze actual samples. Conclusion This strategy has ultrasensitivity and high specificity for DNA methylation quantification, without requiring complex processes such as PCR and enzymatic digestion, which is thus of great value in tumor diagnosis and biomedical research.

scholarly journals Preparation of CdTe/Alginate Textile Fibres with Controllable Fluorescence Emission through a Wet-Spinning Process and Application in the Trace Detection of Hg2+ Ions

Nanomaterials ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 570 ◽  
Author(s):  
Zhihui Zhao ◽  
Cunzhen Geng ◽  
Xihui Zhao ◽  
Zhixin Xue ◽  
Fengyu Quan ◽  
...  
Keyword(s):  
Mechanical Strength ◽  
Resonance Energy Transfer ◽  
Fluorescence Emission ◽  
Resonance Energy ◽  
Coagulation Bath ◽  
Wet Spinning ◽  
Trace Detection ◽  
Metallic Ions ◽  
Textile Fibres ◽  
Cdte Nanocrystals

Fluorescent textile fibres (FTFs) are widely used in many industrial fields. However, in addition to fibres with good fluorescence, fibres with excellent colour controllability, structural stability and appropriate mechanical strength still need to be developed. In this work, CdTe/alginate composite FTFs are prepared by taking advantage of the interactions between CdTe nanocrystals (NCs) and alginate macromolecules via a wet-spinning machine with a CaCl2 aqueous solution as the coagulation bath. CdTe NCs were chemically fixed in the fibre due to the interactions among surface ligands, macromolecules and coagulators (calcium ions), which ensured the excellent dispersity and good stability of the fibres. Förster resonance energy transfer (FRET) between NCs in the fibre was found to be restricted, which means that the emission colour of the fibres was totally controllable and could be predicted. Other properties of alginate fibres, such as flame retardance and mechanical strength, were also well preserved in the fluorescent fibres. Finally, FTFs showed good selectivity toward trace Hg2+ ions over other metallic ions, and the detection could be identified by the naked eye.

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