Direct Label Free Detection of Orchid Virus Using a Micro/Nano Hybrid Structured Biosensor

2019 ◽  
Author(s):  
Wei-Jhen Wang ◽  
Chia-Hwa Lee ◽  
Chin-Wen Li ◽  
Stephen Liao ◽  
Fuh-Jyh Jan ◽  
...  
Keyword(s):  
Label Free ◽  
Self Assembled Monolayer ◽  
Detection Range ◽  
Elisa Kit ◽  
Detection Approach ◽  
Linear Detection ◽  
Label Free Detection ◽  
Semiconductor Fabrication ◽  
Direct Label ◽  
Higher Sensitivity

Abstract In this study, a label-free detection approach for effective detection of the odontoglossum ringspot virus (ORSV) infected orchids has been developed. We used semiconductor fabrication process to fabricate 1,810 micro/nano hybrid structured sensing electrodes on a 8 inch reclaimed wafer. The self-assembled monolayer (SAM) process was then employed to sequentially modify the electrode surface with 11-mercaptoundecanoic acid (11-MUA), 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC)/ N-hydroxysuccinimide (NHS), anti-ORSV, and ORSV. EIS was conducted for the ORSV concentration detection. Experimental results demonstrated that the ORSV concentration in a virus infected orchid leaf could be effectively detected. When compared with the ELISA kit, our device possesses a wider linear detection range (0.5–50,000 ng/mL) and a higher sensitivity. The specificity of our device on ORSV detection was also confirmed. Our sensing device retains advantages, such as label-free, lower amounts of the antibody and target sample required, low detection time, and a wider linear detection range. Those results imply the feasibility of our sensing device in field applications.

scholarly journals Label-Free Electrochemical Biosensor Based on Au@MoS₂–PANI for Escherichia coli Detection

Chemosensors ◽  
2021 ◽  
Vol 9 (3) ◽  
pp. 49
Author(s):  
Pushap Raj ◽  
Man Hwan Oh ◽  
Kyudong Han ◽  
Tae Yoon Lee
Keyword(s):  
Escherichia Coli ◽  
Bacterial Infections ◽  
Limit Of Detection ◽  
Bacterial Pathogens ◽  
Cost Effective ◽  
Covalent Immobilization ◽  
Label Free ◽  
Self Assembled Monolayer ◽  
Detection Range ◽  
Linear Detection

Bacterial infections have become a significant challenge in terms of public health, the food industry, and the environment. Therefore, it is necessary to address these challenges by developing a rapid, cost-effective, and easy-to-use biosensor for early diagnosis of bacterial pathogens. Herein, we developed a simple, label-free, and highly sensitive immunosensor based on electrochemical detection using the Au@MoS₂–PANI nanocomposite. The conductivity of the glassy carbon electrode is greatly enhanced using the Au@MoS₂–PANI nanocomposite and a self-assembled monolayer of mercaptopropionic acid on the gold nanoparticle surface was employed for the covalent immobilization of antibodies to minimize the nonspecific adsorption of bacterial pathogens on the electrode surface. The biosensor established a high selectivity and sensitivity with a low limit of detection of 10 CFU/mL, and detected Escherichia coli within 30 min. Moreover, the developed biosensor demonstrated a good linear detection range, practical utility in urine samples, and electrode regenerative studies.

scholarly journals Comparison of DNAzyme activity for the development of an immobilized heme sensor

MRS Advances ◽  
2018 ◽  
Vol 3 (26) ◽  
pp. 1491-1496
Author(s):  
Natalie Hughes ◽  
Nancy Nguyen ◽  
Deanna-Kaye Daley ◽  
Justin Grennell ◽  
Amira Gee ◽  
...  
Keyword(s):  
Self Assembly ◽  
Point Of Care ◽  
Label Free ◽  
Detection Range ◽  
Chemical Complexity ◽  
Detection Strategy ◽  
Detection Approach ◽  
Label Free Detection ◽  
G Quadruplex ◽  
Care Systems

ABSTRACTPoint-of-care systems require highly sensitive, quantitative and selective detection platforms for the real-time multiplexed monitoring of target analytes. To ensure facile development of a sensor, it is preferable for the detection assay to have minimal chemical complexity, contain no wash steps and provide a wide and easily adaptable detection range for multiple targets. Current studies involve label-free detection strategy for relevant clinical molecules such as heme using G-quadruplex based self-assembly. We have explored the measurement of binding and kinetic parameters of various G-quadruplex/heme complexes which are able to self-associate to form a DNAzyme with peroxidase mimicking capabilities and are critical to nucleic acid research. The detection strategy includes immobilizing the G-quadruplex sequences within a polymer matrix to provide a self-assembly based detection approach for heme that could be translated towards other clinically relevant targets.

scholarly journals Surface Plasmon Resonance Assay for Label-Free and Selective Detection of HIV-1 p24 Protein

Biosensors ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 180
Author(s):  
Lucia Sarcina ◽  
Giuseppe Felice Mangiatordi ◽  
Fabrizio Torricelli ◽  
Paolo Bollella ◽  
Zahra Gounani ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Covalent Binding ◽  
Hiv Infections ◽  
Selectivity Ratio ◽  
Selective Detection ◽  
Label Free ◽  
Self Assembled Monolayer ◽  
Label Free Detection ◽  
P24 Protein ◽  
Hiv 1

The early detection of the human immunodeficiency virus (HIV) is of paramount importance to achieve efficient therapeutic treatment and limit the disease spreading. In this perspective, the assessment of biosensing assay for the HIV-1 p24 capsid protein plays a pivotal role in the timely and selective detection of HIV infections. In this study, multi-parameter-SPR has been used to develop a reliable and label-free detection method for HIV-1 p24 protein. Remarkably, both physical and chemical immobilization of mouse monoclonal antibodies against HIV-1 p24 on the SPR gold detecting surface have been characterized for the first time. The two immobilization techniques returned a capturing antibody surface coverage as high as (7.5 ± 0.3) × 1011 molecule/cm2 and (2.4 ± 0.6) × 1011 molecule/cm2, respectively. However, the covalent binding of the capturing antibodies through a mixed self-assembled monolayer (SAM) of alkanethiols led to a doubling of the p24 binding signal. Moreover, from the modeling of the dose-response curve, an equilibrium dissociation constant KD of 5.30 × 10−9 M was computed for the assay performed on the SAM modified surface compared to a much larger KD of 7.46 × 10−5 M extracted for the physisorbed antibodies. The chemically modified system was also characterized in terms of sensitivity and selectivity, reaching a limit of detection of (4.1 ± 0.5) nM and an unprecedented selectivity ratio of 0.02.

scholarly journals Combination of Capped Gold Nanoslit Array and Electrochemistry for Sensitive Aqueous Mercuric Ions Detection

Nanomaterials ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 88
Author(s):  
Cheng-Chuan Chen ◽  
Shu-Cheng Lo ◽  
Pei-Kuen Wei
Keyword(s):  
Metal Ion ◽  
Water Solution ◽  
Resonant Peak ◽  
Label Free ◽  
Glycerol Water ◽  
Detection Range ◽  
Label Free Detection ◽  
Intensity Changes ◽  
Mercuric Ions ◽  
Nanoslit Arrays

Label-free surface plasmon resonance (SPR) detection of mercuric ions in various aqueous solutions, using capped gold nanoslit arrays combined with electrochemical (EC) sensing technique, is demonstrated. The nanoslit arrays are fabricated on flexible cyclo-olefin polymer substrates by a nanoimprinting lithography method. The EC and SPR signals for the investigation of current responses and transmission SPR spectra are simultaneously measured during metal ions electrodeposition. Glycerol–water solution is studied to evaluate the resonant peak wavelength sensitivity (480.3 nm RIU−1) with a FOM of 40.0 RIU−1 and the obtained intensity sensitivity is 1819.9%. The ferrocyanide/ferricyanide redox couple performs the diffusion controlled electrochemical processes (R2 = 0.99). By investigating the SPR intensity changes and wavelength shifts of various mercuric ion concentrations, the optical properties are evaluated under chronoamperometric conditions. The sensors are evaluated in the detection range between 100 μM and 10 nM with a detection limit of 1 μM. The time dependence of SPR signals and the selectivity of 10 μM Hg2+ in the presence of 10 μM interfering metal ion species from Ca2+, Co2+, Ni2+, Na+, Cu2+, Pb2 + and Mn2+ are determined. The capped gold nanoslit arrays show the selectivity of Hg2+ and the EC sensing method is effectively utilized to aqueous Hg2+ detection. This study provides a label-free detection technique of mercuric ions and this developed system is potentially applicable to detecting chemicals and biomolecules.

scholarly journals An Aptamer-Based Biosensor for Direct, Label-Free Detection of Melamine in Raw Milk

Sensors ◽  
2018 ◽  
Vol 18 (10) ◽  
pp. 3227 ◽  
Author(s):  
Naoto Kaneko ◽  
Katsunori Horii ◽  
Joe Akitomi ◽  
Shintaro Kato ◽  
Ikuo Shiratori ◽  
...  
Keyword(s):  
Sample Preparation ◽  
Background Noise ◽  
Colorimetric Detection ◽  
Raw Milk ◽  
Detection Methods ◽  
Label Free ◽  
Label Free Detection ◽  
High Concentrations ◽  
Direct Label ◽  
Urinary Problems

Melamine, a nitrogen-rich compound, has been used as a food and milk additive to falsely increase the protein content. However, melamine is toxic, and high melamine levels in food or in milk can cause kidney and urinary problems, or even death. Hence, the detection of melamine in food and milk is desirable, for which numerous detection methods have been developed. Several methods have successfully detected melamine in raw milk; however, they require a sample preparation before the analyses. This study aimed to develop an aptamer-DNAzyme conjugated biosensor for label-free detection of melamine, in raw milk, without any sample preparation. An aptamer-DNAzyme conjugated biosensor was developed via screening using microarray analysis to identify the candidate aptamers followed by an optimization, to reduce the background noise and improve the aptamer properties, thereby, enhancing the signal-to-noise (S/N) ratio of the screened biosensor. The developed biosensor was evaluated via colorimetric detection and tested with raw milk without any sample preparation, using N-methylmesoporphyrin IX for fluorescence detection. The biosensor displayed significantly higher signal intensity at 2 mM melamine (S/N ratio, 20.2), which was sufficient to detect melamine at high concentrations, in raw milk.

A nanofluidic device for ultrasensitive and label-free detection of tetracycline in association with γ-cyclodextrin and GO

2021 ◽  
Vol 13 (15) ◽  
pp. 1832-1838
Author(s):  
Jiaxi Cheng ◽  
Fenghua Jiang ◽  
Siqi Zhang
Keyword(s):  
Detection Limit ◽  
Water Samples ◽  
Recovery Rate ◽  
Label Free ◽  
Detection Range ◽  
Low Detection Limit ◽  
Sensing Platform ◽  
Label Free Detection ◽  
Nanofluidic Device

We develop a label-free, sensitive and selective nanochannel sensing platform for detection of TC. The nanosensor provided a low detection limit, a wide detection range and excellent recovery rate in different water samples.

Direct label-free detection of Rotavirus using a hydrogel based nanoporous photonic crystal

RSC Advances ◽  
2016 ◽  
Vol 6 (9) ◽  
pp. 7384-7390 ◽  
Author(s):  
Bohee Maeng ◽  
Youngkyu Park ◽  
Jungyul Park
Keyword(s):  
Photonic Crystal ◽  
Crystal Structures ◽  
Virus Detection ◽  
Label Free ◽  
Label Free Detection ◽  
Near Future ◽  
Direct Label

A direct label-free biosensor based on 3D photonic crystal structures for Rotavirus has been demonstrated. This proposed method will be useful for developing a direct and easy-to-use virus detection kit in the form of POCT in the near future.

scholarly journals An Electrochemical Immunosensor Based on a Self-Assembled Monolayer Modified Electrode for Label-Free Detection of α-Synuclein

Sensors ◽  
2020 ◽  
Vol 20 (3) ◽  
pp. 617 ◽  
Author(s):  
Chuang-Ye Ge ◽  
Md. Mahbubur Rahman ◽  
Wei Zhang ◽  
Nasrin Siraj Lopa ◽  
Lei Jin ◽  
...  
Keyword(s):  
Dynamic Range ◽  
Electrochemical Impedance ◽  
Charge Transfer Resistance ◽  
Label Free ◽  
Serum Samples ◽  
Self Assembled Monolayer ◽  
Electrochemical Impedance Spectra ◽  
Transfer Resistance ◽  
Label Free Detection ◽  
Self Assembled

This research demonstrated the development of a simple, cost-effective, and label-free immunosensor for the detection of α-synuclein (α-Syn) based on a cystamine (CYS) self-assembled monolayer (SAM) decorated fluorine-doped tin oxide (FTO) electrode. CYS-SAM was formed onto the FTO electrode by the adsorption of CYS molecules through the head sulfur groups. The free amine (–NH2) groups at the tail of the CYS-SAM enabled the immobilization of anti-α-Syn-antibody, which concurrently allowed the formation of immunocomplex by covalent bonding with α-Syn-antigen. The variation of the concentrations of the attached α-Syn at the immunosensor probe induced the alternation of the current and the charge transfer resistance (Rct) for the redox response of [Fe(CN)6]3−/4−, which displayed a linear dynamic range from 10 to 1000 ng/mL with a low detection limit (S/N = 3) of ca. 3.62 and 1.13 ng/mL in differential pulse voltammetry (DPV) and electrochemical impedance spectra (EIS) measurements, respectively. The immunosensor displayed good reproducibility, anti-interference ability, and good recoveries of α-Syn detection in diluted human serum samples. The proposed immunosensor is a promising platform to detect α-Syn for the early diagnose of Parkinson’s disease, which can be extended for the determination of other biologically important biomarkers.

scholarly journals Direct and Competitive Optical Grating Immunosensors for Determination of Fusarium Mycotoxin Zearalenone

Toxins ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 43
Author(s):  
Inna Székács ◽  
Nóra Adányi ◽  
István Szendrő ◽  
András Székács
Keyword(s):  
Environmental Safety ◽  
Detection Methods ◽  
Label Free ◽  
Detection Range ◽  
Fusarium Mycotoxin ◽  
Label Free Detection ◽  
Optical Grating ◽  
Covalent Immobilisation ◽  
Magnitude Improvement

Novel optical waveguide lightmode spectroscopy (OWLS)-based immunosensor formats were developed for label-free detection of Fusarium mycotoxin zearalenone (ZON). To achieve low limits of detection (LODs), both immobilised antibody-based (direct) and immobilised antigen-based (competitive) assay setups were applied. Immunoreagents were immobilised on epoxy-, amino-, and carboxyl-functionalised sensor surfaces, and by optimising the immobilisation methods, standard sigmoid curves were obtained in both sensor formats. An outstanding LOD of 0.002 pg/mL was obtained for ZON in the competitive immunosensor setup with a dynamic detection range between 0.01 and 1 pg/mL ZON concentrations, depending on the covalent immobilisation method applied. This corresponds to a five orders of magnitude improvement in detectability of ZON relative to the previously developed enzyme-linked immonosorbent assay (ELISA) method. The selectivity of the immunosensor for ZON was demonstrated with structural analogues (α-zearalenol, α-zearalanol, and β-zearalanol) and structurally unrelated mycotoxins. The method was found to be applicable in maize extract using acetonitrile as the organic solvent, upon a dilution rate of 1:10,000 in buffer. Thus, the OWLS immunosensor method developed appears to be suitable for the quantitative determination of ZON in aqueous medium. The new technique can widen the range of sensoric detection methods of ZON for surveys in food and environmental safety assessment.

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