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1.
ACS Omega ; 9(26): 28648-28658, 2024 Jul 02.
Artículo en Inglés | MEDLINE | ID: mdl-38973936

RESUMEN

Basalt is a suitable target area for CO2 storage. Clarifying the evolution of the pore structure during CO2-fluid-basalt interactions is a crucial element in the preparation of a CO2 geologic storage operation. In this study, a pore-scale phase field model is proposed to simulate the CO2-fluid-basalt interaction process of Snake River Plain basalt. Our model is suitable for complex multimineral natural rocks, and the initial mineral distribution before the reaction can be completely based on a real 3D rock image without the need to simplify the mineral geometry. The simulation results demonstrate that after 120 days of dissolution-precipitation reaction, the volume of secondary minerals (3.75%) exceeds that of dissolved minerals (2.01%), leading to a reduction in porosity by 1.74%. The pore structure of the basalt changes significantly, and the connectivity is obviously reduced. The effects of temperature and pressure on the reaction rate were examined to guide site selection for CO2 sequestration. The results show that increasing temperature and pressure can accelerate the reaction rate, but the impact of pressure on the reaction rate is negligible compared to the significant influence of temperature. Therefore, an area with a high geothermal gradient is conducive to geological sequestration.

2.
Anal Methods ; 13(20): 2313-2319, 2021 05 27.
Artículo en Inglés | MEDLINE | ID: mdl-33956005

RESUMEN

In this work, a sensitive and quantitative immunochromatographic assay (ICA) detection method for avian influenza viruses (AIVs) of the H7 hemagglutinin (HA) antigen was established based on highly chromatic red silica nanoparticles (SiNPs). It can detect two H7 subtypes of influenza viruses, H7N2 and H7N9. The highly chromatic red SiNPs were prepared by adsorbing C.I. Direct Red 224 on the surface of the SiNPs for multiple times using the layer by layer (LbL) self-assembly method under the electrostatic action of ethylene imine polymer (PEI) and poly(sodium-p-styrenesulfonate) (PSS). The highly chromatic red silica nanoparticles modified with anti-H7 HA mAb1 were used as immunodetection probes. The accumulated highly chromatic red SiNPs on the T-line can be observed by the naked eye to qualitatively detect the H7 HA antigen. The quantitative analysis is carried out by using a camera and Image J software. Within the range of 0.1-10 ng mL-1, the linear equation between the H7 HA antigen concentration and the peak area of the T-line gray value was y = 868.9722 + 435.4836X (R2 = 0.9716), and the limit of detection (LOD) of this method was 0.08 pg mL-1 (S/N = 3). The highly chromatic red SiNP based ICA for the detection of H7 HA has no cross activity with other subtypes of influenza viruses. This method of combining highly chromatic colored markers with ICA has great potential in practical applications for the rapid and quantitative detection of other types of AIVs.


Asunto(s)
Subtipo H7N9 del Virus de la Influenza A , Nanopartículas , Animales , Inmunoensayo , Subtipo H7N2 del Virus de la Influenza A , Dióxido de Silicio
3.
Mikrochim Acta ; 187(12): 658, 2020 11 17.
Artículo en Inglés | MEDLINE | ID: mdl-33201356

RESUMEN

Aflatoxin M1 (AFM1) residues in milk pose a major threat to human health, so there is an urgent need for a simple, rapid, and sensitive method for the determination of trace AFM1 in milk. In this study, a competitive immunochromatographic assay (ICA), using visual (PEI/PSS)4 red silica nanoparticles (SiNPs) as signal amplification probes, was used for the highly sensitive detection of AFM1. The (PEI/PSS)4 red SiNPs were used to label AFM1 monoclonal antibody (mAb) to prepare ICA for the detection of AFM1. After exploring the optimal conditions of mAb and immunoprobe dosage conditions, the lowest visual detection limit (VDL) of AFM1 in phosphate-buffered saline with Tween 20 (PBST, 10 mM, pH 7.4, containing 1% BSA, 3% sucrose, 1% trehalose, and 0.5% Tween 20) can reach 0.1 pg/mL. The intuitive visually visible value of AFM1 in both PBST and milk was 10 pg/mL. The results showed that the immunochromatographic system based on high chroma color (PEI/PSS)4 red SiNPs has high sensitivity and broad application prospects for the detection of trace AFM1 residues in milk. The high chroma (PEI/PSS)4 red SiNPs are expected to be a convenient biomarker for improving the sensitivity of immune chromatography bands. Graphical abstract The schematic diagram shows the detection principle. In this work, in the competitive experiment, (PEI/PSS)4 red SiNPs were selected as visual labeling materials, and the specific antibody combined with the labeled material was selected as an immune probe. The AFM1-BSA antigen coupled with the macromolecular BSA was fixed on the T line of the nitrocellulose (NC) membrane. The AFM1 in sample solution competes with AFM1-BSA for the specific binding site of immune probe. The detection sensitivity of this method for AFM1 is obtained by judging the change of the red signal intensity produced by the positive sample, compared with the color at the T line of the negative sample.


Asunto(s)
Aflatoxina M1/análisis , Cromatografía de Afinidad/métodos , Contaminación de Alimentos/análisis , Leche/química , Nanopartículas/química , Polietileneimina/química , Poliestirenos/química , Dióxido de Silicio/química , Aflatoxina M1/inmunología , Animales , Anticuerpos Monoclonales/inmunología , Límite de Detección , Espectroscopía Infrarroja por Transformada de Fourier
4.
Macromol Rapid Commun ; 41(17): e2000313, 2020 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-32767476

RESUMEN

Multiple-stimuli responsive soft actuators with tunable initial shapes would have substantial potential in broad technological applications, ranging from advanced sensors, smart robots to biomedical devices. However, existing soft actuators are often limited to single initial shape and are unable to reversibly reconfigure into desirable shapes, which severely restricts the multifunctions that can be integrated into one actuator. Here, a novel reconfigurable supramolecular polymer/polyethylene terephthalate (PET) bilayer actuator exhibiting multiple-stimuli responses is presented. In this bilayer actuator, the supramolecular polymer layer constructed of poly(5-Norbornene-2-carboxylic acid-1,3-cyclooctadiene) (PNCCO) and azopyridine derivative (PyAzoPy) via H-bonds provides multiple-stimuli responses: PyAzoPy offers light response and carboxylic groups in PNCCO endow the actuator with humidity response. Meanwhile thermoplastic PET layer enables the bilayer actuators to be reconfigured into various shapes by thermal stimuli. The rationally designed actuators exhibit versatile capabilities to reversibly reconfigure into a set of initial shapes and carry out multiple functions, such as photo-driven "foldback-clip" and Ω-shaped crawling robots. In addition, bio-inspired plants constructed by reconfiguration of such actuators demonstrate reversible multiple-stimuli responses. It is anticipated that these novel actuators with highly tunable geometries and actuation modes would be useful to develop multifunctional devices capable of performing diverse tasks.

5.
Mikrochim Acta ; 187(5): 285, 2020 04 23.
Artículo en Inglés | MEDLINE | ID: mdl-32322994

RESUMEN

A competitive immunochromatographic assay (ICA) is presented and used for on-site determination of ractopamine (RAC). Ultramarine blue nanoparticles were directly separated from ultramarine blue industrial products by centrifugation (< 10,000 rpm and > 4000 rpm) and used as visible labels in ICAs. The ultramarine blue nanoparticles were coated by polyacrylic acid (PAA), which provides carboxyl groups on the surface of ultramarine blue nanoparticles. An anti-RAC monoclonal antibody (mAb) was covalently immobilized on the carboxyl-modified ultramarine blue nanoparticle surface via diimide-activated conjugation between the carboxyl groups on the ultramarine blue nanoparticle surface and the amino groups of the antibodies. RAC and BSA-modified RAC competitively bind to the anti-RAC mAb on the ultramarine blue nanoparticles. The blue band in the test line is generated by the accumulation of ultramarine blue nanoparticles and is negatively associated with the RAC content. Under optimal conditions, the visual limit of detection (vLOD) of this ICA for RAC is 2.0 ng mL-1, 2.0 ng mL-1, and 1.0 ng mL-1 in phosphate-buffered saline (PBS), feed samples, and pork samples, respectively. The ultramarine blue nanoparticle-based ICA also shows no cross activity with salbutamol, clorprenaline, clenbuterol, or terbutaline. Graphical abstract Schematic representation of the ultramarine blue nanoparticles immunochromatographic assay for detection of ractopamine (RAC) based on competitive method. The ultramarine blue nanoparticles were screened from commercial ultramarine pigments for the first time and used to detect ractopamine.


Asunto(s)
Colorantes Fluorescentes/química , Nanopartículas/química , Fenetilaminas/análisis , Animales , Anticuerpos Monoclonales/química , Anticuerpos Monoclonales/inmunología , Cromatografía de Afinidad , Estructura Molecular , Tamaño de la Partícula , Fenetilaminas/inmunología , Propiedades de Superficie , Porcinos
6.
Anal Chim Acta ; 1098: 140-147, 2020 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-31948577

RESUMEN

Ultramarine blue particles as a novel visible label has been used to develop immunochromatographic assay (ICA). The ultramarine blue particles, as a sodalite mineral with formula: (Na,Ca)8[(S,Cl,SO4,OH)2(Al6Si6O24)], can generate a blue visible signal were used as a label for ICA. Ultramarine blue particles were applied to a sandwich immunoassay to detect hepatitis B virus surface antigen (HBsAg). Ultramarine blue particles were separated from ultramarine blue industrial product by centrifugation. The polyacrylic acid (PAA) was used to modify the carboxyl group on the surface of ultramarine blue particles. The goat anti-HBsAg monoclonal antibody was modified on ultramarine blue particles by EDC/NHS activation of the carboxyl groups. In the presence of HBsAg, the immune ultramarine blue particles were bound on test line zone and forming a blue line on ICA strip which was directly readout by naked eye and quantitatively measured by Image J software. Under optimal conditions, the color depth of test line was linearly correlated with the concentration of HBsAg in concentration range from 1 to 50 ng mL-1. The calibration equation was y = 385.796 + 97.2298x (R2 = 0.9872), with limit of detection (LOD) of 0.37 ng mL -1(S/N = 3). The sensitivity of this novel ICA was better than that of ICA based on traditional gold nanoparticles as reporter probe. The ultramarine blue particles offer an alternative type of visible label nanomaterial for the development of ICA.


Asunto(s)
Antígenos de Superficie de la Hepatitis B/análisis , Inmunoensayo/métodos , Pigmentos Biológicos/química , Tamaño de la Partícula , Propiedades de Superficie
7.
Mikrochim Acta ; 186(7): 421, 2019 06 11.
Artículo en Inglés | MEDLINE | ID: mdl-31187272

RESUMEN

An immunochromatographic assay (ICA) is presented that can be applied to simultaneous detection of clenbuterol (CLE) and ractopamine (RAC). It is making use of two red and blue silica nanoparticles (SiNPs) that act as labels for encoding the antibodies. This design permits multiplexed analysis in a single test line and does not require an external source for photoexcitation. Anti-CLE was labeled with red SiNPs, and anti-RAC with blue SiNPs. The capture antigens CLE-BSA and RAC-BSA were placed onto the conjugate pad and the test line of the test strip, respectively. Under bare eye examination, no cross-colored lines or nonspecific bioconjugate adsorption were observed, and the visible limit of detections for CLE (red) and RAC (blue) are 3 and 2 ng‧mL-1, respectively. This design allows for multiplexed detection with reduced device dimensions and costs, and with easy integration and manufacturing. Conceivably, the method may be extended to simultaneous determination of numerous other analytes. Graphic abstract The principle of qualitative detection strategy of multiplex immunochromatographic assay for clenbuterol (CLE) and ractopamine (RAC) is schematically illustrated. Depending on the type and ratio of organic dyes, the color of colored silica nanoparticle can be tuned from red to purple and even to black (lower right corner).


Asunto(s)
Agonistas Adrenérgicos beta/análisis , Clenbuterol/análisis , Inmunoensayo/métodos , Nanopartículas/química , Fenetilaminas/análisis , Dióxido de Silicio/química , Agonistas Adrenérgicos beta/inmunología , Anticuerpos Monoclonales/inmunología , Clenbuterol/inmunología , Color , Inmunoensayo/instrumentación , Límite de Detección , Fenetilaminas/inmunología , Pruebas en el Punto de Atención
8.
Anal Chim Acta ; 1038: 97-104, 2018 Dec 14.
Artículo en Inglés | MEDLINE | ID: mdl-30278912

RESUMEN

In this paper, a new type immunochromatographic assay (ICA) based on core-shell red silica nanoparticles (core-shell red SiO2NPs) was proposed and used to detect Escherichia coli O157:H7 (E. coli O157:H7). This is the first report of qualitative ICA for detecting E. coli O157:H7 in phosphate buffer saline (PBS) and food sample using core-shell red SiO2NPs. Monodispersed red SiO2NPs were synthesized in the aqueous solution by modifying amino silane and C.I Reactive Red 136 on unmodified silica nanoparticles. The limit of detection (LOD) of this core-shell red SiO2NPs based ICA for E. coli O157:H7 was 4.5 × 105 CFU/mL in sterile PBS within 20 min. The LOD of this ICA strip for E. coli O157:H7 in milk and pork samples both were 4.5 × 106 CFU/mL. The core-shell red SiO2NPs based ICA for detection of E. coli O157:H7 has no cross activity with other bacteria. All these results show that this new kind of core-shell colored SiO2NPs is promising for the practical applications in ICA and other rapid detection fields.


Asunto(s)
Escherichia coli O157/aislamiento & purificación , Inmunoensayo , Nanopartículas/química , Dióxido de Silicio/química , Tamaño de la Partícula , Dióxido de Silicio/síntesis química
9.
Mikrochim Acta ; 185(10): 455, 2018 09 13.
Artículo en Inglés | MEDLINE | ID: mdl-30215173

RESUMEN

A highly sensitive electrochemical sandwich immunoassay is described for determination of Escherichia coli O157:H7 (E. coli O157:H7). Silica coated magnetite nanoparticles (Fe3O4) were modified with primary antibody to capture E. coli O157:H7. Gold-platinum core/shell nanoparticles (Au@Pt NPs) with different Pt shell thicknesses were prepared via changing the molar ratio of H2PtCl6 to HAuCl4 in the precursor solution. The optimized Au@Pt NPs exhibit enhanced activity in the electrocatalytic reduction of hydrogen peroxide (H2O2). The Au@Pt NPs were modified with graphene that was functionalized with Neutral Red, and then used as an electrochemical label for secondary antibodies and horseradish peroxidase (HRP). The sandwich immunocomplexes were magnetically absorbed on a 4-channel screen printed carbon electrode. Due to the catalysis of the Au@Pt NPs and HRP, the signal is strongly amplified in the presence of H2O2 when using thionine as the electron mediator. Under optimal conditions, the immunoassay has a linear response in the 4.0 × 102 to 4.0 × 108 CFU·mL-1 concentration range, with a limit of detection of 91 CFU·mL-1 (at an S/N ratio of 3). Graphical abstract Preparation of Au@Pt core/shell nanoparticles with different Pt shell thickness (A), rGO-NR (B), rGO-NR-Au@Pt-Ab2-HRP (C) and the preparation and the detection process of the immunoassay (D). rGO: reduced graphene oxide, GO: graphene oxide, NR: Neutral Red, HRP: horseradish peroxidase, AuNPs: gold nanoparticles, Fe3O4@SiO2: Silica coated magnetite nanoparticles, 4-SPCE: 4-channel screen printed carbon electrode.


Asunto(s)
Escherichia coli O157/aislamiento & purificación , Oro/química , Grafito/química , Inmunoensayo/métodos , Nanopartículas de Magnetita/química , Nanopartículas del Metal/química , Platino (Metal)/química , Catálisis , Electroquímica , Peróxido de Hidrógeno/química , Modelos Moleculares , Conformación Molecular , Oxidación-Reducción , Fenotiazinas/química
10.
Anal Biochem ; 559: 34-43, 2018 10 15.
Artículo en Inglés | MEDLINE | ID: mdl-30144412

RESUMEN

Herein, a non-enzymatic sandwich-type electrochemical immunoassay was fabricated for quantitative monitoring of Escherichia coli O157:H7 (E. coli O157:H7). Silica coated Fe3O4 magnetic nanoparticles (Fe3O4@SiO2) were modified with mouse anti-E. coli O157:H7 monoclonal antibody (Ab1) to act as capture probes to reduce detection time and increase the sensitivity of the immunoassay. The Au@Pt nanoparticles were loaded on neutral red (NR) functionalized graphene to form composite complex rGO-NR-Au@Pt. rGO-NR-Au@Pt has high specific surface area and good biocompatibility. rGO-NR-Au@Pt was used as the carriers of detection antibodies (Ab2). Au@Pt catalyzed the reduction of hydrogen peroxide (H2O2) to detection of E. coli O157:H7 with the thionine (TH) as electron mediator to effectually amply the current signal. Under the optimized conditions, a linear relationship between the reduction peak current change (ΔIpc) and the logarithm of the E. coli O157:H7 concentration is obtained in the range from 4.0 × 103 to 4.0 × 108 CFU mL-1 and the limit of detection (LOD) is 4.5 × 102 CFU mL-1 at a signal-to-noise ratio of 3. The immunoassay exhibits acceptable specificity, reproducibility and stability on the detection of E. coli O157:H7. Furthermore, the immunoassay showed good performance in pork and milk samples. The results suggest that this immunoassay will be promising in the food safety area.


Asunto(s)
Técnicas Electroquímicas , Escherichia coli O157/aislamiento & purificación , Oro/química , Grafito/química , Inmunoensayo , Platino (Metal)/química , Tamaño de la Partícula , Propiedades de Superficie
11.
Talanta ; 182: 354-362, 2018 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-29501164

RESUMEN

A sensitive Point-of-Care Testing (POCT) with Au-Pt bimetallic nanoparticles (Au@Pt) functionalized silica nanoparticle (SiO2 NPs) and Fe3O4 magnetic nanoparticles (Fe3O4 NPs) was designed for the quantitative detection of Escherichia coli O157:H7 (E. coli O157:H7). The poly-(4-styrenesulfonic acid-co-maleic acid) (PSSMA) as a negatively charged polyelectrolyte can be easily coated on surface of the amino group modified SiO2 NPs via electrostatic force. PSSMA is also a good stabilizer for water-soluble bimetallic nanostructures. The PSSMA is first time used as a "bridge" to connect the negative charge Au@Pt NPs to the SiO2 NPs, forming Au@Pt/SiO2 NPs. Antibody and invertase conjugated Au@Pt/SiO2 NPs (denoted as Ab/invertase-Au@Pt/SiO2 NPs) were used as signal labels. Monoclonal antibody against E. coli O157:H7 (Ab) functionalized magnetic nanoparticles (denoted as Ab-Fe3O4@SiO2 NPs) were used to enrich and capture the E. coli O157:H7 in positive sample. The immunosensing platform also composed of a personal glucometer (PGM) using for signal readout. Based on this sandwich-type immunoassay, the invertase in the final formed sandwich immunocomplex catalyzed the hydrolysis of sucrose to produce a large amount of glucose for quantitative readout by the PGM. Under optimal conditions, a linear relationship between the glucose concentration and the logarithm of E. coli O157:H7 concentration was obtained in the concentration range from 3.5 × 102 to 3.5 × 108 CFU mL-1 with a detection limit of 1.83 × 102 CFU mL-1 (3σ). This method was used to detect E. coli O157:H7 in spiked milk samples, indicating its potential practical application. This protocol can be applied in various fields of study.


Asunto(s)
Técnicas Biosensibles , Técnicas Electroquímicas , Escherichia coli O157/aislamiento & purificación , Inmunoensayo , Nanopartículas de Magnetita/química , Nanocompuestos/química , Anticuerpos Inmovilizados/química , Anticuerpos Monoclonales/química , Escherichia coli O157/metabolismo , Óxido Ferrosoférrico/química , Glucosa/química , Oro/química , Humanos , Inmunoconjugados/química , Límite de Detección , Nanopartículas de Magnetita/ultraestructura , Maleatos/química , Nanocompuestos/ultraestructura , Sistemas de Atención de Punto , Poliestirenos/química , Dióxido de Silicio/química , Electricidad Estática , Sacarosa/química , beta-Fructofuranosidasa/química
12.
Biosens Bioelectron ; 107: 266-271, 2018 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-29477883

RESUMEN

Here we innovate a portable and quantitative immunochromatographic assay (ICA) with a personal glucose meter (PGM) as readout for the detection of Escherichia coli O157:H7 (E. coli O157:H7). The carboxyl group coated Fe3O4 nanoparticles (MNPs) were synthesized by a one pot method and used as carriers of invertase and monoclonal antibody against E. coli O157:H7. Initially, the invertase and antibody double functionalized MNPs (Invertase-MNPs-IgG) conjugates were prepared and used as label probe in this assay system. Before laminating onto the baking card, the absorbent pad was soaked in sucrose solution and desiccated. MNPs produced brown band at the detection zone of the ICA when acting as direct labels. As they were also coupled with invertase, the invertase catalyzed the hydrolysis of sucrose on the absorbent pad into glucose, which was detected by the PGM. To increase the sensitivity, antibody functionalized MNPs were used to enrich E. coli O157:H7 from sample solution. The innovative aspect of this approach lies in the visualization and quantification of E. coli O157:H7 through Invertase-MNPs-IgG and the detection of glucose concentration using PGM. Although the feasibility is demonstrated using E. coli O157:H7 as a model analyte, this approach can be easily developed to be a universal analysis system and applied to detection of a wide variety of foodborne pathogens and protein biomarkers. This study proposed a qualitative and quantitative analysis device for the clinic diagnostics and food safety analysis.


Asunto(s)
Técnicas Biosensibles/instrumentación , Automonitorización de la Glucosa Sanguínea/instrumentación , Cromatografía de Afinidad/instrumentación , Escherichia coli O157/aislamiento & purificación , Contaminación de Alimentos/análisis , Leche/microbiología , Sistemas de Atención de Punto , Animales , Anticuerpos Inmovilizados/química , Diseño de Equipo , Infecciones por Escherichia coli/microbiología , Análisis de los Alimentos/instrumentación , Glucosa/análisis , Humanos , beta-Fructofuranosidasa/química
13.
Anal Bioanal Chem ; 409(17): 4139-4147, 2017 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-28429065

RESUMEN

In this article, a facile and sensitive electrochemical method for quantification of Salmonella Pullorum and Salmonella Gallinarum (S. Pullorum and S. Gallinarum) was established by monitoring glucose consumption with a personal glucose meter (PGM). Antibody-functionalized magnetic nanoparticles (IgG-MNPs) were used to capture and enrich S. Pullorum and S. Gallinarum, and IgG-MNPs-S. Pullorum and IgG-MNPs-S. Gallinarum complexes were magnetically separated from a sample using a permanent magnet. The trace tag was prepared by loading polyclonal antibodies and high-content glucose oxidase on amino-functionalized silica nanoparticles (IgG-SiNPs-GOx). With a sandwich-type immunoassay format, IgG-SiNPs-GOx were added into the above mixture solution and conjugated to the complexes, forming sandwich composites IgG-MNPs/S. Pullorum and S. Gallinarum/IgG-SiNPs-GOx. The above sandwich composites were dispersed in glucose solution. Before and after the hydrolysis of glucose, the concentration of glucose was measured using PGM. Under optimal conditions, a linear relationship between the decrease of glucose concentration and the logarithm of S. Pullorum and S. Gallinarum concentration was obtained in the concentration range from 1.27 × 102 to 1.27 × 105 CFU mL-1, with a detection limit of 7.2 × 101 CFU mL-1 (S/N = 3). This study provides a portable, low-cost, and quantitative analytical method for bacteria detection; thus, it has a great potential in the prevention of disease caused by S. Pullorum and S. Gallinarum in poultry. Graphical abstract A schematic illustration of the fabrication process of IgG-SiNPs-GOD nanomaterials (A) and IgG-MNPs (B) and experimental procedure of detection of S. Pullorum and S. Gallinarum using GOD-functionalized silica nanospheres as trace tags based on PGM (C).


Asunto(s)
Anticuerpos Inmovilizados/química , Técnicas Electroquímicas/métodos , Glucosa Oxidasa/química , Glucosa/análisis , Nanopartículas/química , Enfermedades de las Aves de Corral/diagnóstico , Salmonelosis Animal/diagnóstico , Salmonella/aislamiento & purificación , Animales , Técnicas Biosensibles/métodos , Pollos/microbiología , Enzimas Inmovilizadas/química , Técnicas para Inmunoenzimas/métodos , Inmunoglobulina G/química , Límite de Detección , Enfermedades de las Aves de Corral/microbiología , Salmonelosis Animal/microbiología , Dióxido de Silicio/química
14.
Mikrochim Acta ; 185(1): 45, 2017 12 09.
Artículo en Inglés | MEDLINE | ID: mdl-29594632

RESUMEN

The authors describe a sandwich-type of electrochemical immunoassay for rapid determination of the foodborne pathogen Cronobacter sakazakii (C. sakazakii). Polyclonal antibody against C. sakazakii (anti-C. sakazakii) and horseradish peroxidase were immobilized on a nanocomposite consisting of reduced graphene oxide, thionine and gold nanoparticles (AuNPs) that was placed on a screen-printed carbon electrode (SPCE). Thionine acts as an electron mediator which also shortens the electron transfer pathway from the conjugated HRP to the electrode surface and amplifies the electrochemical signal. The AuNPs, in turn, improve the electron transfer rate and increase the surface area for capturing antibody. The morphologies of the electrodes were characterized by means of field emission scanning electron microscopy. The electrochemical performance of the immunoassay was evaluated by cyclic voltammetry and differential pulse voltammetry. Under optimal experimental conditions, the electrochemical immunoassay, best operated at a woking potential of -0.18 V (vs. Ag/AgCl) and scan rate of 20 mV/s has a linear response that covers the 8.8 × 104 to 8.8 × 108 CFU·mL-1 C. sakazakii concentration range, with a 1.0 × 104 CFU·mL-1 detection limit (at an S/N ratio of 3). The assay was applied to the detemination of C. sakazakii in spiked infant milk powder and gave recoveries ranging from 92.0 to 105.7%. Graphical abstract A sandwich-type electrochemical immunosensor was designed for C. sakazakii based on the use of rGO. TH, HRP, antibody and AuNPs were anchored on rGO. The nanocomposites were used as traces tag and H2O2 as enzyme substrates. AuNPs were modified on SPCE by electrodeposition.

15.
Anal Chim Acta ; 898: 109-15, 2015 Oct 22.
Artículo en Inglés | MEDLINE | ID: mdl-26526916

RESUMEN

A novel nonenzymatic optical immunoassay strategy was for the first time designed and utilized for sensitive detection of antibody to Salmonella pullorum and Salmonella gallinarum (S. pullorum and S. gallinarum) in serum. The optical immunoassay strategy was based on blue silica nanoparticles (Blue-SiNps) and magnetic beads (MB). To construct such an optical immunoassay system, the Blue-SiNPs were first synthesized by inverse microemulsion method, characterized by SEM, Zeta potential and FTIR. Two nanostructures including Blue-SiNPs and MB were both functionalized with antibody against S. pullorum and S. gallinarum (anti-PG) without using enzyme labeled antibody. Anti-PG functionalized blue silica nanoparticles (IgG-Blue-SiNps) were used as signal transduction labels, while anti-PG functionalized magnetic beads (IgG-MB) were selected to separate and enrich the final sandwich immune complexes. In the process of detecting negative serum, a sandwich immunocomplex is formed between the IgG-MB and IgG-Blue-SiNPs. With the separation of the immunocomplex using an external magnetic field, the final plaque displayed bright blue color. While in the detection of infected serum, IgG-MB and anti-PG formed sandwich immunocomplexes, IgG-Blue-SiNPs were unable to bind to the limited sites of the antigen, and a light brown plaque was displayed in the bottom of microplate well. Stable results were obtained with an incubation time of 60 min at room temperature, and different colors corresponding to different results can be directly detected with naked eye. The reaction of IgG-Blue-SiNPs with S. pullorum was inhibited by 1:100 dilution of positive chicken serum. Such a simple immunoassay holds great potential as sensitive, selective and point-of-care (POC) tool for diagnosis of other biological molecules.


Asunto(s)
Inmunoensayo/métodos , Nanopartículas , Infecciones por Salmonella/diagnóstico , Dióxido de Silicio/química , Microscopía Electrónica de Rastreo
16.
Curr Pharm Biotechnol ; 16(8): 716-23, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-25941889

RESUMEN

Aimed to explore an agglutination test which can simultaneously detect two pathogenic bacteria, an agglutination test based on colored silica nanoparticles (colored-SiNps) was established in this work. Monodisperse colored-SiNps were used as agglutination test carriers; red-SiNps and blue-SiNps were prepared by reverse microemulsion with C.I. Reactive red 136 and C.I. Reactive Blue 14. Then the red-SiNps were sensitized with antibodies against E. sakazaki and denoted as IgG-red-SiNps; The blue-SiNps were coated with antibodies against S. pullorum and S. Gallinarum and denoted as IgGblue- SiNps. The mixture solution of IgG-red-SiNps and IgG-blue-SiNps could simultaneously agglutinate with E. sakazakii and S. pullorum and S. gallinarum on glass slide. The E. sakazakii and S. pullorum and S. gallinarum could be simultaneously detected by agglutination test with obvious agglutination phenomena. The E. sakazakii and S. pullorum and S. gallinarum could both be detected in a range from 4×10(3) to 4×10(9) CFU/mL. The pullorum and S. gallinarum and E. sakazakii in the infected food sample were detected by mixture solution of IgG-red-SiNps and IgG-blue-SiNps too. This agglutination test was easy and rapid, it might be useful for in situ rapid detection method for simultaneously screening different pathogenic microorganisms of foods and feeds in the field.


Asunto(s)
Pruebas de Aglutinación/métodos , Enterobacter/aislamiento & purificación , Nanopartículas , Salmonella/aislamiento & purificación , Dióxido de Silicio/química , Pruebas de Aglutinación/instrumentación , Microscopía Electroquímica de Rastreo , Nanopartículas/química , Nanopartículas/ultraestructura
17.
J Microbiol Methods ; 106: 110-118, 2014 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-25193438

RESUMEN

In order to increase the reproducibility and stability of electrochemical immunosensor, which is a key issue for its application and popularization, an accurate and stable immunosensor for rapid detection of Salmonella pullorum (S. pullorum) was proposed in this study. The immunosensor was fabricated by modifying Screen-printed Carbon Electrode (SPCE) with electrodeposited gold nanoparticles (AuNPs), HRP-labeled anti-S. pullorum and ionic liquids (ILs) (AuNP/HRP/IL). AuNPs are electrodeposited on the working electrode surface to increase the amount of antibodies that bind to the electrode and then modified with ILs to protect the antibodies from being inactivated in the test environment and maintain their biological activity and the stability of the detection electrode. The electrochemical characteristics of the stepwise modified electrodes and the detection of S. pullorum were investigated by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). As shown in the results of the experiments, AuNPs with unique electrochemical properties as well as biocompatibility characteristics have been proven to be able to strengthen the antibody combination effectively and to increase the electrochemical response signal. In addition, a crucial assessment regarding implementation of stability and reproducibility analysis of a range of immunosensors is provided. We found that application of AuNPs/ILs in the immune modified electrodes showed obvious improvement when compared with other groups. Given their high levels of reproducibility, stability, target specificity and sensitivity, AuNPs and ILs were considered to be excellent elements for electrode modification.


Asunto(s)
Técnicas Biosensibles/métodos , Galvanoplastia , Oro/análisis , Líquidos Iónicos , Nanopartículas/análisis , Salmonella/aislamiento & purificación , Anticuerpos Antibacterianos/inmunología , Reproducibilidad de los Resultados , Salmonella/inmunología , Sensibilidad y Especificidad
18.
Anal Biochem ; 434(2): 218-20, 2013 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-23219558

RESUMEN

A disposable immunosensor based on electrochemically reduced graphene oxide (ERGO) was developed for the detection of Enterobacter sakazakii. First, the graphene was deposited on a screen-printed carbon electrode (SPCE) by an electrochemical method. Second, the horseradish peroxidase-labeled bacteria-specific antibody was assembled onto the modified electrode to enhance the sensitivity of the immunosensor. The immunosensor constructed in this study can sensitively and rapidly detect E. sakazakii, and this method may contribute to further foodborne pathogen detection. In addition, this ERGO-modified SPCE could also provide new insights into the development of immunosensors for other bacteria to facilitate rapid detection.


Asunto(s)
Cronobacter sakazakii , Técnicas Electroquímicas , Grafito/química , Inmunoensayo/instrumentación , Óxidos/química , Equipos Desechables , Electrodos , Microscopía de Fuerza Atómica , Oxidación-Reducción
19.
Wei Sheng Wu Xue Bao ; 51(4): 538-46, 2011 Apr.
Artículo en Chino | MEDLINE | ID: mdl-21796990

RESUMEN

OBJECTIVE: To explore a new rapid detection method for detecting of Food pathogens. METHOD: We used the Smartongue, to determine the composition informations of the liquid culture samples and combined with soft independent modelling of class analogies (SIMCA) to analyze their respective species, then set up a Smartongue -SIMCA model to discriminate the V. parahaemolyticus. RESULTS: The Smartongue has 6 working electrodes and three frequency segments, we can built 18 discrimination models in one detection. After comparing all the 18 discrimination models, the optimal working electrodes and frequency segments were selected out, they were: palladium electrode in 1 Hz frequency segment, tungsten electrode in 100 Hz and silver electrode in 100 Hz. Then 10 species of pathogenic Vibrio were discriminated by the 3 models. The V. damsela, V. metschnikovii, V. alginalyticus, V. cincinnatiensis, V. metschnikovii and V. cholerae O serogroup samples could be discriminated by the SIMCA model of V. parahaemolyticus with palladium electrode 1 Hz frequency segment; V. mimicus and V. vulnincus samples could be discriminated by the SIMCA model of V. parahaemolyticus with tungsten electrode 100 Hz frequency segment; V. carcariae and V. cholerae non-O serogroup samples could be discriminated with the SIMCA model of V. parahaemolyticus in silver electrode 100 Hz frequency segment. The accurate discrimination of ten species of Vibrio samples is 100%. CONCLUSION: The Smartongue combined with SIMCA can discriminate V. parahaemolyticus with other pathogenic Vibrio effectively. It has a promising future as a new rapid detection method for V. parahaemolyticus.


Asunto(s)
Técnicas Biosensibles/métodos , Microbiología de Alimentos/métodos , Vibrio parahaemolyticus/clasificación , Vibrio parahaemolyticus/aislamiento & purificación , Vibrio parahaemolyticus/química
20.
Anal Chim Acta ; 690(2): 240-7, 2011 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-21435482

RESUMEN

A novel voltammetric electronic tongue, smart tongue, was employed to monitor the growth of mold and to differentiate between four types of mold grown in liquid medium. Principal component analysis (PCA) was used to extract the relevant information obtained by the smart tongue. Reference growth curves were based on measurements of dry weight and pH. The growth detected by the smart tongue was basically consistent with that observed by the measurement of dry weight and pH. The optimal combinations of electrodes and frequencies for monitoring growth were as follows: for Aspergillus, both the Pt and Au electrodes at 1 Hz, 10 Hz and 100 Hz; for Penicillium, the Pt and W electrodes at 100 Hz; for Mucor, the Pt, Pd and W electrodes at the three frequency segments; for Rhizopus, the Pd, Ti and Ag electrodes at the three frequency segments. The Ag electrode at 10 Hz or 100 Hz frequency could differentiate well between the four types of mold for culturing 6 h in the liquid media. Therefore, the smart tongue has a promising future as a modern rapid analytical technology for the real time detection of the growth of mold and for the classification model of mold.


Asunto(s)
Medios de Cultivo/química , Técnicas Electroquímicas/métodos , Electrónica , Hongos/aislamiento & purificación , Aspergillus/crecimiento & desarrollo , Aspergillus/aislamiento & purificación , Técnicas Biosensibles/métodos , Técnicas Electroquímicas/instrumentación , Electrodos , Hongos/crecimiento & desarrollo , Oro/química , Concentración de Iones de Hidrógeno , Plomo/química , Mucor/crecimiento & desarrollo , Mucor/aislamiento & purificación , Penicillium/crecimiento & desarrollo , Penicillium/aislamiento & purificación , Platino (Metal)/química , Análisis de Componente Principal , Rhizopus/crecimiento & desarrollo , Rhizopus/aislamiento & purificación , Plata/química , Tungsteno/química
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