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1.
A highly effective strategy for quantification of plasmid which was a special dsDNA based on bisPNA by electrochemical impedance spectroscopy was presented in this work. Firstly, through Au?S bond, thiol‐terminated bisPNA probes were immobilized onto the gold electrode surface. Then bisPNA probes directly hybridized with target plasmid DNA pBR322 based on the PNA.DNA‐PNA invasion triplex without denaturation. In the presence of redox electroactive ions [Fe(CN)6]3?/4? as hybridization indicator, the charge transfer resistance (Rct) was produced, and Rct was measured via electrochemical impedance spectroscopy. Under optimal conditions, this strategy showed a good linear relationship between the ΔRct which was the difference of Rct obtained before and after bisPNA hybridized with plasmid pBR322, and logarithm of the concentration of plasmid pBR322 within the range from 1 nM to 100 nM (R2=0.993), with a limit of detection (LOD) of 0.1 nM. Furthermore, this bisPNA‐assisted biosensor showed good stability and satisfactory analytical reliability. In addition, this novel bisPNA‐assisted biosensor also exhibited excellent analytical results in human serum.  相似文献   

2.
An early on time detection of breast cancer significantly affects the treatment process and outcome. Herein, a new label-free impedimetric biosensor is developed to determine the lowest change in the level of calreticulin (CALR), which is a new biomarker of breast carcinoma. The proposed immunosensor is fabricated by using reduced graphene oxide/amino substituted polypyrrole polymer (rGO-PPyNH2) nanocomposite modified disposable electrode. The anti-CALR antibodies are first attached on the rGO-PPyNH2 nanocomposite coated electrode through glutaraldehyde crosslinking; the CALR antigens are then immobilized with the addition of CALR antigens to form an immunocomplex on the sensing surface. This immunocomplex induces considerably larger interfacial electron transport resistance (Rct). The variation in the Rct has a linear relationship with CALR level in the detection range of 0.025 to 75 pg mL−1, with a detection limit of 10.4 fg mL−1. The suggested biosensor shows high selectivity to CALR, good storage stability (at least 5 weeks) and suitable reproducibility results as shown in quality control chart. The designed immunosensor is utilized to analyze CALR levels in human sera with satisfying results. This immunosensor provides a novel way for the clinical determination of CALR and other cancer biological markers.  相似文献   

3.
《Electroanalysis》2017,29(5):1267-1277
Graphite rod (GR) modified with electrochemicaly deposited gold nanoparticles (AuNPs) and adsorbed glucose oxidase (GOx) was used in amperometric glucose biosensor design. Enzymatic formation of polypyrrole (Ppy) on the surface of GOx/AuNPs/GR electrode was applied in order to improve analytical characteristics and stability of developed biosensor. The linear glucose detection range for Ppy/GOx/AuNPs/GR electrode was dependent on the duration of Ppy‐layer formation and the linear interval was extended up to 19.9 mmol L−1 after 21 h lasting synthesis of Ppy. The sensitivity of the developed biosensor was determined as 21.7 μA mM−1 cm−2, the limit of detection – 0.20 mmol L−1. Ppy/GOx/AuNPs/GR electrodes demonstrated advanced good stability (the t 1/2 was 9.8 days), quick detection of glucose (within 5 s) in the wide linear interval. Additionally, formed Ppy layer decreased the influence of electroactive species on the analytical signal. Developed biosensor is suitable for the determination of glucose in human serum samples.  相似文献   

4.
《Electroanalysis》2018,30(5):910-920
A label‐free DNA biosensor based on magnetite/multiwalled carbon nanotubes/chitosan (Fe3O4/MWCNTs‐COOH/CS) nanomaterial for detection of Bacillus cereus DNA sequences was fabricated. Negatively charged DNA was electrostatically adsorbed onto materials by protonation of positively charged chitosan under acidic conditions. The electrode surface and hybridization process were carried out by cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS). Under optimal conditions, the biosensor showed a good linear relationship between peak currents difference (ΔI) and logarithm of the target DNA concentration (Log C) ranging from 2.0×10−13 to 2.0×10−6 M with a detection limit of 2.0×10−15 M (signal/noise ratio of 3). The biosensor also revealed an excellent selectivity to three‐base, completely mismatched and completely matched DNA. This is a simple, fast and friendly method with a low detection limit for the detection of Bacillus cereus specific DNA compared with previously reported electrochemical DNA biosensor. Furthermore, the DNA biosensor may lead to the development of a technology for gold prospecting in the wild.  相似文献   

5.
Bismuth film modified and chemically activated carbon micro‐thread electrodes were investigated for the simultaneous determination of Cd(II) and Pb(II) using square wave anodic stripping voltammetry. The carbon thread electrode was characterised using both surface and electrochemical techniques. Electrochemical impedance spectroscopy (EIS) studies demonstrated that the H2SO4/IPA‐treated carbon thread electrode showed a much improved resistance response (Rct=23 Ω) compared to the IPA‐untreated carbon thread (Rct=8317 Ω). Furthermore, parameters such as the effect of deposition potential, deposition time and Bi(III) concentration were explored using square wave voltammetry. Detection limits (S/N=3) for Cd(II) and Pb(II) were found to be 1.08 µg L?1 and 0.87 µg L?1, respectively and response was found to be linear over the range 5–110 µg L?1. The proposed Bi/IPA‐treated carbon thread electrode exhibited a high selectivity towards Cd(II) and Pb(II) even in the presence of a range of heavy metals and is capable of repetitive and reproducible measurements, being attributed to the high surface area, geometry and electrode treatment characteristics. The proposed metal ion sensor was employed to determine cadmium and lead in river water samples and % RSD was found to be 5.46 % and 5.93 % for Cd(II) and Pb(II) respectively (n=3). Such facile sensing components favour the development of cost effective portable devices for environmental sample analysis and electrochemical applications.  相似文献   

6.
《Electroanalysis》2018,30(8):1659-1668
PAMAM dendrimer/reduced graphene oxide nanocomposite modified pencil graphite electrode (PAMAM/RGO/PGE) was used to fabricate an electrochemical DNA biosensor for determination of Rituxan (RTX) at low concentrations, for the first time. The fabricated biosensor was characterized with FE‐SEM, EIS, and CV techniques. The ds‐DNA/PAMAM/RGO/PGE was used as a working electrode to study the interaction between the RTX and salmon sperm ds‐DNA by DPV technique. Because of the interaction between the drug and DNA leads to a decrease in the guanine oxidation peak current, it was used as an indicator for the determination of the RTX. Under the optimized experimental conditions, a wide linear relationship between RTX concentration and guanine signal was obtained within the range of 7.0 to 60.0 μmol L−1 and 60.0 to 300.0 μmol L−1 with a low detection limit (0.56 μmol L−1). To clarify the interaction mechanism between the RTX and the ds‐DNA, DPV and UV‐Vis measurements were used. The reproducibility, stability, and performance of the constructed biosensor was examined by quantitative measuring RTX in pharmaceutical and human serum samples with good precision (RSD; 2.0–6.0 %) and acceptable recoveries (100.04–101.95 %).  相似文献   

7.
The preparation and characterisation of a new composite electrode with Co3O4 particles-modified multi-walled carbon nanotube (MWCNT) and poly(phenosafranine), as well as its novel application for the voltammetric detection of rutin was described. The resulting composite electrode was characterised using scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and electrochemical impedance spectroscopy (EIS). In the optimised experimental conditions, the oxidation peak current (Ipa) of rutin showed a linear increase in concentration, between 0.008–0.6 and 0.80–6.0 μmol L−1, with a detection limit of 0.00379 μmol L−1. Due to its good selectivity and stability, the composite electrode was successfully applied in detecting rutin in pharmaceutical formulations.  相似文献   

8.
《Electroanalysis》2017,29(12):2698-2707
A cholesterol biosensor based on cholesterol oxidase‐poly(diallyldimethylammonium chloride)‐carbon nanotubes‐nickel ferrite nanoparticles (ChOx‐PDDA‐CNTs‐NiFe2O4NPs) solution is easily fabricated by using a single dropping step on a glassy carbon electrode (GCE) surface. This technique is an alternative way to reduce complexity, cost and time to produce the biosensor. The uniformly dispersed materials on the electrode surface enhance the catalytic reaction of cholesterol oxidase and electron transfer from the oxidation of hydrogen peroxide in the system. The nickel ferrite nanoparticles were synthesized by co‐precipitation and calcination at various temperatures. These nanoparticles were then characterized using field emission scanning electron microscopy (FE‐SEM), energy‐dispersive X‐ray spectroscopy (EDX), Fourier transform infrared spectroscopy (FTIR), cyclic voltammetry (CV) and X‐ray diffraction (XRD). The synthesized material calcined at 700 °C was well defined and presented the octahedral metal stretching with cubic NiFe2O4NPs phase. In cyclic voltammetric study, the ChOx‐PDDA‐CNTs‐NiFe2O4NPs/GCE showed 0.43 s−1 charge transfer rate constant (K s), 7.79×10−6 cm2 s−1 diffusion coefficient value (D ), 0.13 mm2 electroactive surface area (A e) and 3.58×10−8 mol cm−2 surface concentration ( ). This modified electrode exhibits stability in term of percent relative standard deviation (%RSD=0.62 %, n=10), reproducibility (%RSD=0.81, n=10), high sensitivity (25.76 nA per mg L−1 cm−2), linearity from 1 to 5,000 mg L−1 (R2=0.998) with a low detection limit (0.50 mg L−1). Its Michaelis‐Menten constant (K m) was 0.14 mM with 0.92 μA maximum current (I max) and demonstrated good selectivity without the effects of electroactive species such as ascorbic acid, glucose and uric acid. The cholesterol biosensor was successfully applied to determine cholesterol levels in human blood samples, showing promise due to its simplicity and availability.  相似文献   

9.
《Electroanalysis》2017,29(11):2665-2671
Detection of Enterotoxigenic Escherichia coli in various biological samples has tremendous importance in human health. In this direction, we have designed a label free electrochemical biosensor for highly selective detection of Escherichia coli through detecting ST gene. The ability of sensor probe to detect STG was confirmed using polymerase chain reaction. The biosensor was fabricated based on STG specific probes immobilized on platinum nanoparticles chitosan nanocomposite on screen printed carbon electrode, which was characterized by cyclic voltammetry, transmission electron microscopy, and fourier transform infrared spectroscopy. A highly sensitive label free sensing was achieved by analyzing STG hybridization using electrochemical impedance spectroscopy (EIS) technique. The EIS analysis showed a significant increase in charge transfer resistance after STG interaction with the highly selective ssDNA probe immobilized on the nanocomposite film. The increase in charge transfer resistance was evaluated for varying concentrations of STG, which shows a dynamic range between 1.0×10−12 and 1.0×10−4 with the detection limit of 3.6×10−14 M (RSD<4.5 %). The regeneration of sensor probe was also studied and interference due to non‐target sequences was evaluated to ensure the selectivity of the designed sensor. The practical applicability of sensor probe was also analyzed by detecting the STG from the bacteria present in surface water.  相似文献   

10.

For the first time, the synergistic effect of graphene oxide nanocolloids (nano-GO) and silicon dioxide (silica) nanoparticles (SiO2-nanoparicles) has been used to modify a glassy carbon electrode (GCE) for the determination of gallic acid (GA). The modified electrode surface was characterised by using scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDXA) and Fourier transform infrared spectroscopy (FTIR). The electrochemical behaviour of the modified electrode was then studied, using cyclic voltammetry (CV) and differential pulse voltammetry (DPV), showing that the electrode was sensitive to GA in a concentration range of 6.25 × 10−6 to 1.0 × 10−3 mol L−1, with a correlation coefficient R2 of 0.9956 and a limit of detection of 2.09 × 10−6 mol L−1 (S/N = 3). The proposed method was successfully used for the determination of GA in red wine, white wine and orange juice, with recoveries of 102.3, 95.4 and 97.6%, respectively.

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11.
A label‐free DNA biosensor based on three‐dimensional reduced graphene oxide (3D‐rGO) and polyaniline (PANI) nanofibers modified glassy carbon electrode (GCE) was successfully developed for supersensitive detection of breast cancer BRCA1. The results demonstrated that 3D‐rGO and PANI nanofibers had synergic effects for reducing the charge transfer resistance (Rct), meaning a huge enhancement in electrochemical activity of 3D‐rGO‐PANI/GCE. Probe DNA could be immobilized on 3D‐rGO‐PANI/GCE for special and sensitive recognition of target DNA (1.0×10?15–1.0×10?7 M) with a theoretical LOD of 3.01×10?16 M (3S/m). Furthermore, this proposed nano‐biosensor could directly detect BRCA1 in real blood samples.  相似文献   

12.
A method for Ti/TiO2 photoelectrode preparation using laser calcination instead of oven calcination process was introduced. The prepared TiO2 film was investigated by X-ray diffraction (XRD), electrochemical impedance spectroscopy (EIS) and amperometry, and it was found that the prepared electrode mainly consisted of anatase TiO2 nanoparticles on its surface and exhibited a superior photocatalytic activity. The electrode was employed as a sensor to measure chemical oxygen demand (COD) of the wastewater. The measuring principle was based on the photocurrent responses of the electrode which were proportional to the COD values. Under the optimized experimental conditions, the linear range was 50–2000 mg L−1, and the detection limit was 16 mg L−1 (S/N=3). This method was characterized by short analysis time, simplicity, low environmental impact and long lifetime of the sensor. Additionally, the COD values obtained from the proposed and conventional methods agreed well as demonstrated by the high significant correlation between the two sets of COD values (R=0.9895, n=25).  相似文献   

13.
Development of an electrochemical DNA biosensor for the direct detection and discrimination of double-stranded oligonucleotide (dsDNA) corresponding to hepatitis C virus genotype 3a, without its denaturation, using a gold electrode is described. The electrochemical DNA sensor relies on the modification of the gold electrode with 6-mercapto-1-hexanol and a self-assembled monolayer of 14-mer peptide nucleic acid probe, related to the hepatitis C virus genotype 3a core/E1 region. The increase of differential pulse voltammetric responses of methylene blue, upon hybridization of the self-assembled probe with the target ds-DNA to form a triplex is the principle behind the detection and discrimination. Some hybridization experiments with non-complementary oligonucleotides were carried out to assess whether the developed DNA sensor responds selectively to the ds-DNA target. Diagnostic performance of the biosensor is described and the detection limit was found to be 1.8 × 10−12 M in phosphate buffer solution, pH 7.0. The relative standard deviation of measurements of 100 pM of target ds-DNA performed with three independent probe-modified electrodes was 3.1%, indicating a remarkable reproducibility of the detection method.  相似文献   

14.
In this work, a renewable tyrosinase-based biosensor was developed for the detection of catechol, using a carbon black paste electrode, without any mediator. The effect of pH, type of electrolyte, and amount of tyrosinase enzyme were explored for optimum analytical performance. The best-performing biosensor in amperometric experiments at potential −0.2 V vs. Ag/AgCl (3 mol L−1 KCl) was obtained using a 0.1 mol L−1 phosphate buffer solution (pH 7.0) as electrolyte. Under optimized conditions, the proposed biosensor had two concentration linear ranges from 5.0×10−9 to 4.8×10−8 and from 4.8×10−8 to 8.5×10−6 mol L−1 and a limit of detection of 1.5×10−9 mol L−1. The apparent Michaelis-Menten constant ( ) was calculated by the amperometric method, and the obtained value was 1.2×10−5 mol L−1 whose result was similar when compared with other studies previously. The biosensor was applied in river water samples, and the results were very satisfactory, with recoveries near 100 %. In addition, the response of this biosensor for different compounds, taking into account their molecular structures was investigated and the results obtained showed no interference with the response potential of catechol. The electrochemical biosensor developed in this work can be considered highly advantageous because it does not require the use of a mediator (direct detection) for electrochemical response, and also because it is based on a low-cost materials that can be used with success to immobilise other enzymes and/or biomolecules.  相似文献   

15.
In this paper, we report a new PNA biosensor for electrochemical detection of point mutation or single nucleotide polymorphism (SNP) in p53 gene corresponding oligonucleotide based on PNA/ds-DNA triplex formation following hybridization of PNA probe with double-stranded DNA (ds-DNA) sample without denaturing the ds-DNA into single-stranded DNA (ss-DNA). As p53 gene is mutated in many human tumors, this research is useful for cancer therapy and genomic study. In this approach, methylene blue (MB) is used for electrochemical signal generation and the interaction between MB and oligonucleotides is studied by differential pulse voltammety (DPV). Probe-modified electrode is prepared by self-assembled monolayer (SAM) formation of thiolated PNA molecules on the surface of Au electrode. A significant increase in the reduction signal of MB following hybridization of the probe with the complementary double-stranded oligonucleotide (ds-oligonucleotide) confirms the function of the biosensor. The selectivity of the PNA sensor is investigated by non-complementary ds-oligonucleotides and the results support the ability of the sensor to detect single-base mismatch directly on ds-oligonucleotide. The influence of probe and ds-DNA concentrations on the effective discrimination against complementary sequence and point mutation is studied and the concentration of 10?6 M is selected as appropriate concentration. Diagnostic performance of the biosensor is described and the detection limit is found to be 4.15 × 10?12 M.  相似文献   

16.
An enzymatic biosensor has been developed for the determination of selenate (SeO42−), in which selenate reductase (SeR) is chemically attached to a gold disk electrode by lipoic acid N-hydroxysuccinimide ester as linker, allowing the catalytic reduction of the SeO42− to SeO32−. Modification of the gold electrode was characterized by X-ray photoelectron spectroscopy (XPS), time-of-flight secondary ion mass spectroscopy (ToF-SIMS), and electrochemistry. Cyclic voltammetry (CV) and differential pulse voltammetry (DPV) measurements were performed in different buffers for selenate determination. Under optimum conditions, the calibration curve was linear over the range 7.0–3900.0 μg L−1 with limits of detection and quantification of 4.97 and 15.56 μg L−1, respectively. The possible interference of the relevant oxyanions SO42−, NO3, NO2, PO43− and AsO43− in the determination of SeO42− was studied. Finally, the proposed biosensor was used to determine SeO42− with recovery between 95.2 and 102.4 % in different real water samples.  相似文献   

17.
18.
A novel electrochemical sensor based on Cu-MOF-199 [Cu-MOF-199 = Cu3(BTC)2 (BTC = 1,3,5-benzenetricarboxylicacid)] and SWCNTs (single-walled carbon nanotubes) was fabricated for the simultaneous determination of hydroquinone (HQ) and catechol (CT). The modification procedure was carried out through casting SWCNTs on the bare glassy carbon electrode (GCE) and followed by the electrodeposition of Cu-MOF-199 on the SWCNTs modified electrode. Cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and scanning electron microscopy (SEM) were performed to characterize the electrochemical performance and surface characteristics of the as-prepared sensor. The composite electrode exhibited an excellent electrocatalytic activity with increased electrochemical signals towards the oxidation of HQ and CT, owing to the synergistic effect of SWCNTs and Cu-MOF-199. Under the optimized condition, the linear response range were from 0.1 to 1453 μmol L−1 (RHQ = 0.9999) for HQ and 0.1–1150 μmol L−1 (RCT = 0.9990) for CT. The detection limits for HQ and CT were as low as 0.08 and 0.1 μmol L−1, respectively. Moreover, the modified electrode presented the good reproducibility and the excellent anti-interference performance. The analytical performance of the developed sensor for the simultaneous detection of HQ and CT had been evaluated in practical samples with satisfying results.  相似文献   

19.
A new strategy was introduced for ssDNA immobilization on a modified glassy carbon electrode. The electrode surface was modified using polyaniline and chemically reduced graphene oxide decorated cerium oxide nanoparticles (CeO2NPs-RGO). A single-stranded DNA (ssDNA) probe was immobilized on the modified electrode surface. Fast Fourier transform square wave voltammetry (FFT-SWV) was applied as detection technique and [Ru(bpy)3]2+/3+ redox signal was used as electrochemical marker. The hybridization of ssDNA with its complementary target caused a dramatic decrease in [Ru(bpy)3]2+/3+ FFT-SW signal. The proposed electrochemical biosensor was able to detect Aeromonas hydrophila DNA oligonucleotide sequence encoding aerolysin protein. Under optimal conditions, the biosensor showed excellent selectivity toward complementary sequence in comparison with noncomplementary and two-base mismatch sequences. The dynamic linear range of this electrochemical DNA biosensor for detecting 20-mer oligonucleotide sequence of A. hydrophila was from 1 × 10−15 to 1 × 10−8 mol L−1. The proposed biosensor was successfully applied for the detection of DNA extracted from A. hydrophila in fish pond water up to 0.01 μg mL−1 with RSD of 5%. Besides, molecular docking was applied to consider the [Ru(bpy)3]2+/3+ interaction with ssDNA before and after hybridization.  相似文献   

20.
In this study; a sensitive, selective, and simple electrochemical sensor was developed to determine low concentration pyridoxine (Py) using a phosphorus-doped pencil graphite electrode (P-doped/PGE). Electrode modification was implemented using the chronoamperometry method at +2.0 V constant potential and 100 seconds in 0.1 mol L−1 H3PO4 supporting electrolyte solution. The characterization processes of the P-doped/PGE were carried out using electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), X-ray photoelectron spectroscopy (XPS), field emission scanning electron microscopy (FESEM), energy-dispersive X-ray spectroscopy (EDX), and atomic force microscope (AFM) methods. In the concentration study, using the differential pulse voltammetry (DPV) method, a linear calibration plot was acquired in the concentration range of 0.5 to 300 μmol L−1 Py. The limit of quantification (LOQ) and limit of detection (LOD) of the developed method were calculated as 0.219 μmol L−1 and 0.0656 μmol L−1, respectively. Detection of Py has been successfully performed on the P-doped/PGE in the beverage samples. As a result, the method developed has been shown to have fast, low cost, and simple for the sensitive and selective detection of Py as an effective electrode.  相似文献   

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