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The presence of sulfamethoxazole (SMX) in natural waters has become a significant concern recently because of its detrimental effects on human health and the ecological environment. To address this issue, it is of utmost urgency to develop a reliable method that can determine SMX at ultra-low levels. In our research, we utilized PVP-induced shape control of a hydrothermal synthesis method to fabricate layer-like structured VS2, and employed it as an electrode modification material to prepare an electrochemical sensor for the sensitive determination of SMX. Thus, our prepared VS2 electrodes exhibited a linear range of 0.06-10.0 µM and a limit of detection (LOD) as low as 47.0 nM (S/N = 3) towards SMX detection. Additionally, the electrochemical sensor presented good agreement with the HPLC method, and afforded perfect recovery results (97.4-106.8%) in the practical analysis. The results validated the detection accuracy of VS2 electrodes, and demonstrated their successful applicability toward the sensitive determination of SMX in natural waters. In conclusion, this research provides a promising approach for the development of electrochemical sensors based on VS2 composite materials.
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Antibacterianos , Compostos de Vanádio , Humanos , Sulfametoxazol , Técnicas Eletroquímicas/métodos , Eletrodos , Limite de DetecçãoRESUMO
Photochemical (PEC) sensors were severely limited for multiplex detection applications due to the cross interference between multiplex signals at the single recognition interface. In this work, a distance-regulated PEC sensor was developed for multiplex detection by using an i-Motif sequence with conformational transformation activity as the signal transduction unit. Through dynamic regulation of the spatial distance between the end site of the functional sequence and the electrode material, the photogenerated electrons on the surface of the sensor were directionally transferred. Thus, a PEC sensor with "signal-on" and "signal-off" dual signal output modes was developed for simultaneous detection of multitarget molecules. Combining isothermal nucleic acid amplification, the PEC sensor constructed in this work was successfully applied to the detection of two virus (Norovirus and Rotavirus) nucleic acid sequences. Under the optimal condition, this bioassay protocol exhibits a linear range of 0.01-100 nM for both viruses with detection limits of 0.72 and 0.53 pM, respectively. In this study, a stimulus-mediated distance regulation strategy successfully addressed the transduction of multiplex detection signals at the single recognition interface of the PEC sensor. It is expected that the technical barriers to multiplex detection of PEC sensors will be overcome and the application of PEC sensing technology will be expanded in the field of environmental analysis.
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Norovirus , Ácidos Nucleicos , Bioensaio , Eletrodos , ElétronsRESUMO
The plasmonic colorimetric biosensors are very favorable for the on-site testing and naked-eye screening of analytes from real samples, but how to realize their highly sensitive assays with simple manipulations is still a great challenge. Herein we designed a target-triggered dual cascade nucleic acid recycling strategy to amplify the assembly of a hyperbranched DNA nanostructure and thus developed a novel kanamycin colorimetric biosensing method. The first cycle arising from the aptamer recognition-triggered strand displacement reaction and its cascade cycle constructed on the catalytic reaction of two nucleases could release an output DNA to trigger the assembly of the DNA nanostructure. Based on the high capture of alkaline phosphatase at this DNA nanostructure to induce the localized surface plasmon resonance change of gold nanobipyramids (Au NBPs), an ultrasensitive colorimetric signal transduction strategy was constructed. Through the measurement of the shift of the characteristic absorption wavelength of Au NBPs, a very wide linear range from 10 fg mL-1 to 1 ng mL-1 and a very low detection limit of 1.4 fg mL-1 were obtained. Meanwhile, the obvious multicolor change of Au NBPs could be used for the visual semi-quantitative analysis of Kana residues. The whole homogeneous assay process well simplified the manipulation and also ensured the excellent repeatability. These excellent performances determine the great potential of the method for future applications.
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Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Nanopartículas Metálicas , Nanoestruturas , Ácidos Nucleicos , Colorimetria/métodos , Limite de Detecção , DNA/genética , DNA/química , Técnicas Biossensoriais/métodos , Ouro/química , Aptâmeros de Nucleotídeos/química , Nanopartículas Metálicas/químicaRESUMO
Nitrite is both an environmental contaminant and a food additive. Excessive intake of nitrites not only causes blood diseases, but also has the potential risk of causing cancer. Therefore, rapid detection of nitrite in water is necessary. In this work, we propose an electrochemical sensor for the sensing of nitrite. Glassy carbon electrodes modified with noble metal nanomaterials have been widely used in the preparation of sensors, but the surface properties of noble metals largely affect the sensing performance. This work proposes the biosynthesis of Au nanoparticles using the pollen extract of Lycoris radiata as a reducing agent. Flavonoids rich in pollen can be used as weak reducing agents for the reduction of chloroauric acid, and slowly synthesize uniformly dispersed Au nanoparticles. These Au nanoparticles do not agglomerate because they contain small biological molecules on the surface and can form a homogeneous sensing interface on the electrode surface. The electrochemical sensor assembled with biosynthesized Au nanoparticles provides linear detection of nitrite between 0.01 and 3.8 mM. The sensor also has excellent immunity to interference. In addition, the proposed sensor was also successfully used for the detection of nitrite in drinking water.
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Água Potável , Nanopartículas Metálicas , Água Potável/análise , Técnicas Eletroquímicas , Ouro/química , Nanopartículas Metálicas/química , NitritosRESUMO
By utilizing a target biorecognition reaction to induce the self-assembly of G-quadruplexes and the aggregation of gold nanoparticles (Au NPs), this work develops a novel colorimetric biosensing method for kanamycin (Kana) antibiotic detection. The compact G-quadruplex structure was assembled from its two half-split sequences which were designed in two hairpin substrates of the Mg2+-dependent DNAzyme (MNAzyme). Besides hybridizing with the aptamer strand, the MNAzyme sequence was also split into two half fragments to be designed in the two substrates. Upon the aptamer-recognition reaction toward Kana, the MNAzyme strand could be quantitatively released to cause the exposure of the split G-quadruplex-sequences on two hairpin substrate-modified Au NPs and simultaneous release of two half fragments of the MNAzyme-sequence. Thus, the K+-assisted self-folding of G-quadruplexes causes the cross-linking of the two Au NPs to realize the Au NP aggregation-based colorimetric signal output (measured at the largest absorption peak near 520 nm). Meanwhile, the self-assembled formation of the second MNAzyme drastically amplified the signal response. Under the optimal conditions, a wide linear range from 0.1 pg mL-1 to 10 ng mL-1 and an ultrahigh sensitivity with the detection limit of 76 fg mL-1 were obtained. The dose-recovery experiments in real samples showed satisfactory results with recoveries from 98.4 to 105.4% and relative errors compared with the ELISA method less than 4.1%. Due to the high selectivity, excellent repeatability and stability, and simple manipulation, this method indicates a promising potential for practical applications. A novel homogeneous biosensing method was developed for the convenient detection of the kanamycin antibiotic. The target biorecognition-induced and dual DNAzyme-catalytic assembly of G-quadruplexes enabled the amplified aggregation of gold nanoparticles for the simple, cheap, stable, and ultrasensitive colorimetric signal transduction of the method.
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DNA Catalítico , Quadruplex G , Nanopartículas Metálicas , Antibacterianos , DNA Catalítico/química , Ouro/química , Canamicina , Nanopartículas Metálicas/química , OligonucleotídeosRESUMO
Herein, we report a Fe3O4@polydopamine (PDA) nanocomposite and exonuclease III (Exo III)-assisted homogeneous fluorescence biosensing method for ultrasensitive detection of kanamycin (Kana) antibiotic. A hairpin DNA containing the Kana-aptamer sequence (HP) was first designed for the highly specific biorecognition of the target analyte. Because of the aptamer biorecognition-induced structural change of HP and the highly effective catalyzed reaction of Exo III, a large amount of fluorophore labels were released from the designed fluorescence DNA probe. During the homogeneous reaction process, the Exo III-assisted dual recycling significantly amplified the fluorescence signal output. Moreover, the excessive probes were easily adsorbed and separated by the Fe3O4@PDA nanocomposite, which decreased the background signal and increased the signal-to-noise ratio. These strategies result in the excellent analytical performance of the method, including a very low detection limit of 0.023 pg mL-1 and a very wide linear range of six orders of magnitude. In addition, this method has convenient operation, excellent selectivity, repeatability and satisfactory reliability, and does not involve the design and utilization of complicated DNA sequences. Thus, it exhibits a promising prospect for practical applications.
Assuntos
Técnicas Biossensoriais , Canamicina , Antibacterianos , Exodesoxirribonucleases , Indóis , Limite de Detecção , Polímeros , Reprodutibilidade dos TestesRESUMO
Chloramphenicol (CAP) is a harmful compound associated with human hematopathy and neuritis, which was widely used as a broad-spectrum antibacterial agent in agriculture and aquaculture. Therefore, it is significant to detect CAP in aquatic environments. In this work, carbon nanotubes/silver nanowires (CNTs/AgNWs) composite electrodes were fabricated as the CAP sensor. Distinguished from in situ growing or chemical bonding noble metal nanomaterials on carbon, this CNTs/AgNWs composite was formed by simple solution blending. It was demonstrated that CNTs and AgNWs both contributed to the redox reaction of CAP in dynamics, and AgNWs was beneficial in thermodynamics as well. The proposed electrochemical sensor displayed a low detection limit of up to 0.08 µM and broad linear range of 0.1-100 µM for CAP. In addition, the CNTs/AgNWs electrodes exhibited good performance characteristics of repeatability and reproducibility, and proved suitable for CAP analysis in real water samples.
Assuntos
Cloranfenicol , Nanotubos de Carbono , Nanofios , Cloranfenicol/análise , Técnicas Eletroquímicas , Eletrodos , Humanos , Reprodutibilidade dos Testes , PrataRESUMO
When the target biorecognition-triggered assembly of two Mg2+-dependent DNAzymes (MNAzymes) is employed for dually catalytic release of peroxidase-mimicking G-quadruplex DNAzymes (G-DNAzymes), this work develops a novel homogeneous colorimetric method for an ultrasensitive bioassay of platelet-derived growth factor-BB (PDGF-BB). The first MNAzyme assembly is realized through a highly specific aptamer biorecognition-driven proximity ligation reaction. Its catalytic cleavage toward the two designed hairpin substrates not only releases a large amount of G-DNAzymes for colorimetric signal transduction but also enables the spontaneous assembly of another MNAzyme for signal amplification. This leads to the successful detection of PDGF-BB in a wide linear range from 2.0 pg mL-1 to 20 ng mL-1 with a very low detection down to 0.088 pg mL-1. As the whole reactions including aptamer biorecognitions, DNA hybridizations, and catalytic cleavages of MNAzymes are conducted in a homogeneous solution, this method has very simple manipulations and also has high repeatability. In addition, the high specificity of the aptamer biorecognition-triggered signal transduction decides the excellent selectivity of the method. This bioassay does not require an expensive instrument and nucleic acid labeling for signal readout or any nanomaterial, enzyme, or nuclease for signal amplification. Thus, it displays an extensive potential for clinical diagnostic applications.
Assuntos
DNA Catalítico/metabolismo , Magnésio/metabolismo , Fator de Crescimento Derivado de Plaquetas/análise , Biocatálise , Colorimetria , Quadruplex G , Humanos , Fator de Crescimento Derivado de Plaquetas/metabolismoRESUMO
Herein we combine the sandwich immunoreaction at a vertically aligned single-walled carbon nanotube (SWCNT)-based immunosensor and the enzymatically catalytic deposition of gold nanoparticles (Au NPs) by a gold nanoprobe to develop a novel electrochemical immunosensing method. The vertically arranged nanostructure was prepared through the covalent linking of terminally carboxylated SWCNTs at an aryldiazonium-modified electrode. It not only provides an excellent platform for the high density immobilization of antibodies to obtain the immunosensor but also serves as useful molecular wires to accelerate electron transfer during the electrochemical immunosensing process. Meanwhile, the enzymatic reaction of the nanoprobe prepared by surface functionalization of the nanocarrier of Au NPs by high-content glucoamylases can catalyze the deposition of a large number of Au NPs at the immunosensor. The electrochemical stripping analysis of these nanoparticles enabled the convenient signal transduction of the method. Due to the sensitive gold stripping analysis at the vertically aligned SWCNTs and the multi-enzyme signal amplification of the nanoprobe, the electrochemical signal response was greatly enhanced. Thus, the method can be used for the ultrasensitive detection of the tumor biomarker of carcinoembryonic antigen in a wide linear range of 5 orders of magnitude with a low detection limit of 0.48 pg mL-1. Considering its obvious performance superiorities, this immunosensing method exhibits an extensive prospect for practical applications.
Assuntos
Biomarcadores Tumorais/sangue , Antígeno Carcinoembrionário/sangue , Técnicas Eletroquímicas/métodos , Imunoensaio/métodos , Nanopartículas Metálicas/química , Nanotubos de Carbono/química , Anticorpos Imobilizados/imunologia , Aspergillus niger/enzimologia , Biomarcadores Tumorais/imunologia , Antígeno Carcinoembrionário/imunologia , Técnicas Eletroquímicas/instrumentação , Eletrodos , Glucana 1,4-alfa-Glucosidase/química , Ouro/química , Humanos , Limite de DetecçãoRESUMO
A magnetic bead (MB)-based sandwich biorecognition reactions is combined with a gold nanoprobe-induced homogenous synthesis of molybdophosphate to develop a novel bioassay method for the electrochemical detection of the tumor biomarker of carcinoembryonic antigen (CEA). The nanoprobe is prepared through the specific loading of numerous alkaline phosphatase (ALP)-functionalized gold nanoparticles (Au NPs) on a double-stranded DNA (dsDNA) produced by the CEA aptamer-triggered hybridization chain reaction (HCR). Both the large amounts of PO43- produced by the ALP catalytic hydrolysis of pyrophosphate and the phosphate backbones of dsDNA can react with the added MoO42- to generate electroactive molybdophosphates. So, the gold nanoprobe was used for signal tracing of the sandwich bioassay of CEA at a constructed antibody-functionalized MB platform. The sensitive electrochemical measurement of molybdophosphate produced from the quantitatively captured nanoprobes at a carbon nanotube-modified electrode (measured at about 0.12 V vs. Ag/AgCl, 3 M KCl) enabled the convenient signal transduction of the method. Due to the dually enhanced synthesis of molybdophosphate by the HCR and multi-enzyme Au NP nanotags, this method shows a wide linear range from 0.05 pg mL-1 to 10 ng mL-1 along with a low detection limit of 0.027 pg mL-1. In addition, the MB-based biorecognition reaction and the homogeneous synthesis of molybdophosphate are much convenient in manipulations. These excellent performances decide the extensive application potentials of the method. Graphical abstract A magnetic bead-based bioassay method was simply developed for the electrochemical detection of carcinoembryonic antigen. The dually enhanced homogenous synthesis of molybdophosphate by hybridization chain reaction (HCR) and enzyme nanotags and the sensitive electrochemical measurement of molybdophosphate at a carbon nanotube (CNT)-electrode enable ultrasensitive signal transduction of the method.
Assuntos
Fosfatase Alcalina/química , Antígeno Carcinoembrionário/sangue , Técnicas Eletroquímicas/métodos , Molibdênio/química , Ácidos Fosfóricos/química , Anticorpos Imobilizados/imunologia , Aptâmeros de Nucleotídeos/química , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/química , Antígeno Carcinoembrionário/química , Antígeno Carcinoembrionário/imunologia , DNA/química , Enzimas Imobilizadas/química , Ouro/química , Humanos , Imunoensaio/métodos , Separação Imunomagnética , Limite de Detecção , Nanopartículas Metálicas/química , Técnicas de Amplificação de Ácido Nucleico , Ácidos Fosfóricos/síntese químicaRESUMO
In this work, a kind of environment-friendly and water-dispersible silicon nanodot (SiND) was rapidly synthesized by using the mild reagents (3-aminopropyl)triethoxysilane (APTES) and glucose. It was found that the fluorescence of the as-prepared SiNDs can be quenched obviously by permanganate due to the inner filter effect. Inspired by this finding, a novel fluorescent sensor for sensitive detection of hydrogen peroxide (H2O2) was developed through the oxidation-reduction reaction between permanganate and H2O2. The detection limit of H2O2 is down to 2.8 nM. Since H2O2 is an important molecule and involved in various studies, this sensor could be applied in various H2O2-related biological analyses. As a proof-of-application demonstration, a sensitive biosensor for glucose detection was constructed through the catalytic oxidation of glucose to generate H2O2. The as-constructed sensor showed good linear response to glucose over the range from 0.16 to 16 µM with a detection limit of 0.11 µM. Moreover, the biosensor can be readily extended to other sensors for different targets, which indicates the broad applications of the proposed sensing strategy in biomedical analysis.
RESUMO
By employment of an aptamer-initiated hybridization chain reaction (HCR) to enhance the enzyme biomineralization of cupric subcarbonate, this work develops a novel colorimetric biosensing method for protein analysis. The HCR product was used to specifically attach a large amount of urease-functionalized gold nanoparticles (Au NPs) for the preparation of a gold nanoprobe. After the sandwich biorecognition reactions, this nanoprobe could be quantitatively captured onto the antibody-functionalized magnetic bead (MB) platform. Then, numerous copper ions would be enriched onto the MB surface through the urease-induced biomineralization of cupric subcarbonate. Based on the complete release of Cu2+ ions for the sensitive copper chromogenic reaction, convenient colorimetric signal transduction was thus achieved for the quantitative analysis of the target analyte of the carcinoembryonic antigen. The HCR product provides a large number of biotin sites for the attachment of Au NP nanotags. The biomineralization reaction of high-content urease loaded onto Au NPs leads to highly efficient Cu2+ enrichment for signal amplification. So this method features excellent performance including a very wide linear range and a low detection limit down to 0.071 pg mL-1. In addition, the satisfactory results of real sample experiments reveal that this method possesses huge potential for practical applications.
Assuntos
Biomarcadores Tumorais/sangue , Técnicas Biossensoriais/métodos , Antígeno Carcinoembrionário/sangue , Animais , Anticorpos Imobilizados/imunologia , Anticorpos Monoclonais/imunologia , Aptâmeros de Nucleotídeos/química , Biomarcadores Tumorais/imunologia , Biomineralização , Carbonatos/química , Antígeno Carcinoembrionário/imunologia , Colorimetria/métodos , Cobre/química , Ouro/química , Humanos , Imunoensaio/métodos , Separação Imunomagnética , Limite de Detecção , Nanopartículas Metálicas/química , Coelhos , Urease/químicaRESUMO
The authors of "A glassy carbon electrode modified with N-doped carbon dots for improved detection of hydrogen peroxide and paracetamol (Microchimica Acta 185, no. 2 (2018): 87)" wish to replace the incorrect images of Fig. 1C, 1D shown below.
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The authors describe a nanocomposite that was obtained by in-situ deposition of CdS nanocrystals on mesoporous silica nanospheres (MSNs), and its use in an electrochemical immunoassay of human immunoglobulin G (HIgG). The MCN/CdS nanocomposite was covalently modified with the antibodies against HIgG and then employed in a voltammetric immunoassay at antibody-functionalized magnetic beads. Through sandwich immunoreaction, the MCN/CdS nanoprobes are quantitatively captured onto the magnetic beads where numerous Cd(II) ions are released in an acidic solution. The Cd(II) can be detected by anodic stripping voltammetry at a typical working potential of -0.78 V (vs. Ag/AgCl). In combination with the high loading of CdS on MSNs, the use of the stripping voltammetric analysis renders the method high sensitivity. A wide linear range varying from 0.01 to 100 ng mL-1 is obtained for HIgG detection with a lower detection limit at 2.9 pg mL-1. In addition, the preparation of the nanoprobe is inexpensive. The magnetic bead-based assay does not require complex manipulations. Therefore, this method is deemed to possess a wide scope in that it may be applied to other immunoassays. Graphical abstract Graphical Abstract contains poor quality and small text inside the artwork. Please do not re-use the file that we have rejected or attempt to increase its resolution and re-save. It is originally poor, therefore, increasing the resolution will not solve the quality problem. We suggest that you provide us the original format. We prefer replacement figures containing vector/editable objects rather than embedded images. Preferred file formats are eps, ai, tiff and pdf.A TIFF file at 900 dpi resolution of the Graphical Abstract has been attached via this online system. Schematic presentation of the preparation of the mesoporous silica nanosphere (MSN)/CdS nanocomposite for the electrochemical immunoassay of human IgG at magnetic beads. The high decoration of CdS on MSN and the stripping voltammetric analysis of Cd(II) ions render the method high sensitivity.
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Cádmio/análise , Imunoensaio/métodos , Imunoglobulina G/análise , Compostos de Cádmio/química , Técnicas Eletroquímicas , Humanos , Nanopartículas/química , Nanosferas/química , Dióxido de Silício/química , Sulfetos/químicaRESUMO
Nitrogen doped carbon dots (NCDs) were synthesized using a low temperature approach and used to modify a glassy carbon electrode (GCE) via dipping. The oxygen groups on the surface of the NCDs, and the charge delocalization of the NCDs warrant an excellent electrocatalytic activity of the GCE toward oxidation of paracetamol (PA) and reduction of H2O2. PA and H2O2 were detected at 0.34 V and -0.4 V (both vs. Ag/AgCl) using differential pulse voltammetry and amperometric I-T measurement, respectively. The modified GCE has a linear response to PA in the 0.5 to 600 µM concentration range, and to H2O2 in the 0.05 µM to 2.25 mM concentration range. The detection limits are 157 nM and 41 nM, respectively. In our perception, the modified GCE holds promise for stable, selective and sensitive determination of PA and H2O2 in pharmaceutical analysis. Graphic abstract Nitrogen doped carbon dots (NCDs) were synthesized and used to modify a glassy carbon electrode. Surface functional groups on NCDs can trigger electrocatalytic reactions toward paracetamol oxidation and H2O2 reduction with high sensitivities.
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Ferrocene-grafted dendrimer was covalently linked to the surface of a carbon nanotubes (CNTs)-chitosan (CS) nanocomposite modified electrode for immobilizing high-content glucose oxidase (GOx), which resulted in the successful development a novel reagentless glucose biosensor. Electrochemical impedance spectroscopy, cyclic voltammetry, and amperometry were used to characterize the preparation process and the enzymatically catalytic response of this biosensor. Due to the excellent electron transfer acceleration of the CNTs and the high-content loading of the GOx biomolecule and ferrocene mediator on the electrode matrix, this biosensor showed excellent analytical performance such as fast response time less than 10 s, wide linear range from 0.02 to 2.91 mM and low detection limit down to 7.5 µM as well as satisfactory stability and reproducibility toward the amperometric glucose determination. In addition, satisfactory result was obtained when it was used for the glucose measurements in human blood samples. Thus this biosensor provides great potentials for practical applications.
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Técnicas Biossensoriais/métodos , Glicemia/análise , Enzimas Imobilizadas/metabolismo , Glucose Oxidase/metabolismo , Nanotubos de Carbono/química , Dendrímeros/química , Técnicas Eletroquímicas/métodos , Eletrodos , Enzimas Imobilizadas/química , Compostos Ferrosos/química , Glucose Oxidase/química , Humanos , Limite de Detecção , Metalocenos/química , Reprodutibilidade dos TestesRESUMO
A nanoporous gold nanosphere (pAu NS) was synthesized to load high-content glucose oxidase (GOx) and ferrocene (Fc) for the successful preparation of a new gold nanoprobe. After the specific recognition of the tumor biomarker of carcinoembryonic antigen (CEA) at a gold electrode based aptasensor, this GOx and Fc dually functionalized pAu NS nanoprobe was further used for sandwich immunoreaction and signal tracing. Based on the Fc-mediated GOx-catalytic reaction, the gold nanoprobes quantitatively captured onto the electrode surface produced a sensitive electrochemical signal corresponding to the protein recognition events, which led to the development of a new biosensing method for CEA measurement. Both the high loading of GOx and Fc on the pAu NS nanocarrier and the enzymatically catalytic reaction of the nanoprobe greatly amplify the electrochemical signal; meanwhile, the immobilization of the Fc mediator on this enzyme nanoprobe and the highly specific aptamer recognition drastically decrease the background current, resulting in the achievement of ultrahigh sensitivity of the method. Under optimum conditions, this method shows an excellent analytical performance including a wide linear relationship of five-order of magnitude and a low detection limit down to 0.45 pg mL(-1). Thus this pAu NS based gold nanoprobe and the proposed immunoassay method provide great potential for practical applications.
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Biomarcadores Tumorais/análise , Técnicas Biossensoriais , Antígeno Carcinoembrionário/análise , Glucose Oxidase/química , Nanopartículas Metálicas , Metalocenos/química , Técnicas Eletroquímicas , Ouro , Humanos , Imunoensaio , Limite de Detecção , NanoporosRESUMO
A novel ultrasensitive immunoassay method was developed based on the electrochemical measurement of polyaniline, which was catalytically produced by horseradish peroxidase-functionalized gold nanoparticle (HRP-Au NP) probe at an immunosensor. The immunosensor was prepared step-wise by first modifying the electrode with reduced graphene oxide (rGO)/Au NPs nanocomposite followed by the immobilization of capture antibodies on its surface. After performing a sandwich immunoreaction, the quantitatively captured HRP-Au NP nanoprobes could catalyze oxidation of aniline to produce electroactive polyaniline on the immunosensor surface. The electrochemical measurement of polyaniline enabled a novel detection strategy for HRP-based immunoassay. Both the signal amplification of the HRP-Au NP nanoprobe and the electron transfer acceleration of rGO/Au NPs on the immunosensor surface greatly improved the detection sensitivity of the immunoassay method. With the use of human IgG as a model analyte, this method showed a wide linear range over 4 orders of magnitude with a detection limit of 9.7 pg/mL. In addition, the immunosensor had low cost, satisfactory reproducibility and stability, and acceptable reliability. The relatively positive potential range for the polyaniline measurement completely excluded the conventional interference from dissolved oxygen. Thus, this method provides a promising potential for practical applications.
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Compostos de Anilina/metabolismo , Técnicas Biossensoriais/métodos , Peroxidase do Rábano Silvestre/metabolismo , Imunoensaio/métodos , Animais , Eletroquímica , Ouro/química , Grafite/química , Peroxidase do Rábano Silvestre/química , Humanos , Nanopartículas Metálicas/química , Óxidos/química , Reprodutibilidade dos Testes , Propriedades de SuperfícieRESUMO
Photoelectrochemical (PEC) biosensors have exhibited a promising potential for assays of a large variety of analytes; however, how to realize their low background-based "super on" signal output is still a great challenge. Herein, we report a novel multiple nucleic acid amplification-assisted "on-off-super on" signal transition mechanism for the PEC biosensing of kanamycin antibiotics. The biosensing platform was constructed on a perylene-3,4,9,10-tetracarboxylic dianhydride-based photoelectrode, and its strong photocurrent could be well inhibited by an anchored ferrocene (Fc)-labeled hairpin DNA to produce a low background signal. Two target biorecognition-triggered exonuclease III-catalytic reactions were adopted to produce an annular four-footed DNA walker (AFW) and a methylene blue (MB)-labeled DNA strand. By using their synergistic effect to release Fc quenchers and simultaneously capture MB sensitizers, a "super on" signal output was realized. As a result, a very wide linear range from 10 fg mL-1 to 10 ng mL-1 and an ultra-low detection limit of 7.8 fg mL-1 were obtained. Meanwhile, the aptamer recognition-based homogeneous reaction and AFW-based multiple nucleic acid amplification effectively simplified the assay manipulation and well ensured the repeatability of the method. The satisfactory sample experiment results indicated its good reliability and accuracy for the antibiotic residue analysis application.