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1.
J Dairy Sci ; 105(3): 2108-2118, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-34998563

RESUMEN

Foods contaminated by foodborne pathogens have always been a great threat to human life. Herein, we constructed an electrochemical immunosensor for Salmonella detection by using a Fe3O4@graphene modified electrode. Because of the excellent electrical conductivity and mechanical stability of graphene and the large specific surface area of Fe3O4, the Fe3O4@graphene nanocomposite exhibits an excellent electrical signal, which greatly increased the sensitivity of the immunosensor. Gold nanoparticles were deposited on Fe3O4@graphene nanocomposite by electrochemical technology for the immobilization of the antibody. Cyclic voltammetry was selected to electrochemically characterize the construction process of immunosensors. The microstructure and morphology of related nanocomposites were analyzed by scanning electron microscopy. Under optimized experimental conditions, a good linear relationship was achieved in the Salmonella concentration range of 2.4 × 102 to 2.4 × 107 cfu/mL, and the limit of detection of the immunosensor was 2.4 × 102 cfu/mL. Additionally, the constructed immunosensor exhibited acceptable selectivity, reproducibility, and stability and provides a new reference for detecting pathogenic bacteria in milk.


Asunto(s)
Técnicas Biosensibles , Grafito , Nanopartículas del Metal , Nanocompuestos , Animales , Técnicas Biosensibles/veterinaria , Carbono , Técnicas Electroquímicas/veterinaria , Electrodos , Oro/química , Grafito/química , Inmunoensayo/veterinaria , Límite de Detección , Nanopartículas del Metal/química , Leche , Nanocompuestos/química , Reproducibilidad de los Resultados , Salmonella
2.
J Dairy Sci ; 105(4): 2895-2907, 2022 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-35181133

RESUMEN

Salmonella, as a common foodborne pathogen in dairy products, poses a great threat to human health. We studied a new detection method based on quantum dots (QD). A fluorescent biosensor with multiple fluorescent signal amplification based on a streptavidin (SA) biotin system and the polyamino linear polymer poly-l-lysine (PLL) were established to detect Salmonella in milk. First, Salmonella was captured on a black 96-well plate with paired Salmonella mAb to form a double-antibody sandwich. Second, SA was immobilized on biotin-modified mAb by SA-biotin specific bond. Then, the biotin-modified polylysine (BT-PLL) was bound on SA and specifically bonded again through the SA-biotin system. Finally, water-soluble CdSe/ZnS QD-labeled SA was added to a black 96-well plate for covalent coupling with BT-PLL. The fluorescent signal was amplified in a dendritic manner by the layer-by-layer overlap of SA and biotin and the covalent coupling of biotinylated PLL. Under optimal conditions, the detection limit was 4.9 × 103 cfu/mL in PBS. The detection limit was 10 times better than that of the conventional sandwich ELISA. In addition, the proposed biosensor was well specific and could be used for detecting Salmonella in milk samples.


Asunto(s)
Técnicas Biosensibles , Puntos Cuánticos , Animales , Técnicas Biosensibles/veterinaria , Biotina/química , Leche , Polilisina , Salmonella , Estreptavidina/química
3.
J Dairy Sci ; 104(2): 1494-1503, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33246628

RESUMEN

Rapid and sensitive detection technology is the key to preventing food-borne disease outbreaks. In this study, a low-field nuclear magnetic resonance (NMR) biosensor based on polyamidoamine dendrimers was prepared for the rapid detection of Salmonella in milk. The polyamidoamine dendrimer was biotinylated by amide reaction and chelated to diethylene triamine pentacetate acid and gadolinium to form magnetic complexes. The antibody and magnetic complexes were combined through a streptavidin-biotin system using streptavidin as an intermediate bridge to obtain the immunoprobe. Salmonella was captured by the immunoprobe via antigen-antibody interaction and then separated from the mixture by membrane filtration. Finally, the longitudinal relaxation signal of the filtrate was obtained by NMR. The biosensor had excellent anti-interference capability and could detect Salmonella within 1.5 h at a sensitivity of 103 cfu mL-1. This method based on NMR can realize detection in complex samples and has the potential to be a quick and nondestructive method for detecting target bacteria.


Asunto(s)
Técnicas Biosensibles , Enfermedades Transmitidas por los Alimentos/microbiología , Gadolinio/química , Leche/microbiología , Poliaminas/química , Salmonella/aislamiento & purificación , Animales , Reacciones Antígeno-Anticuerpo , Biotina/química , Dendrímeros/química , Femenino , Filtración , Enfermedades Transmitidas por los Alimentos/prevención & control , Espectroscopía de Resonancia Magnética , Sensibilidad y Especificidad , Espectroscopía Infrarroja por Transformada de Fourier , Estreptavidina/química , Factores de Tiempo
4.
J Dairy Sci ; 104(11): 11486-11498, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-34454766

RESUMEN

Rapid and sensitive detection of foodborne pathogens is of great importance for food safety. Here, a set of nuclear magnetic resonance (NMR) biosensors based on a O-carboxymethyl chitosan target gadolinium (Gd) probe was developed to quickly detect Salmonella in milk by combining NMR technology and bioimmunotechnology with membrane filtration technology. First, O-carboxymethyl chitosan (O-CMC) was biotinylated to prepare biotinylated O-carboxymethyl chitosan (biotin-O-CMC) through amide reaction, and biotinylated magnetic complexes (biotin-O-CMC-Gd) were obtained by using O-CMC, which has strong chelating adsorption on Gd. The target probe was obtained by combining biotin-O-CMC-Gd with the biotinylated antibody (biotin-antibody) via streptavidin (SA) by introducing the SA-biotin system. Then, Salmonella was captured by the target probe through antigen-antibody interaction. Finally, NMR was used to measure the longitudinal relaxation time (T1) of the filtrate collected by membrane filtration. This NMR biosensor with good specificity and high efficiency can detect Salmonella with the sensitivity of 1.8 × 103 cfu/mL within 2 h; in addition, it can realize the detection of complex samples because of its strong anti-interference capability and may open up a new method for rapid detection of Salmonella, which has a great application potential.


Asunto(s)
Técnicas Biosensibles , Biotina , Animales , Técnicas Biosensibles/veterinaria , Quitosano/análogos & derivados , Gadolinio , Espectroscopía de Resonancia Magnética , Leche , Salmonella , Estreptavidina
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