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A rapid NMR method for the detection of foodborne pathogens based on indirect enrichment of paramagnetic nano-Fe probes

A food-borne pathogenic bacteria, paramagnetic nanotechnology, used in measurement devices, analysis by nuclear magnetic resonance, instruments, etc.

Inactive Publication Date: 2016-01-20
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no literature reporting this method at home and abroad.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0020] Examine food samples to determine whether they contain the harmful pathogen Listeria monocytogenes.

[0021] 1. Preparation of nano-Fe immunoprobes: Antibody 1 was rabbit anti-IgG monoclonal antibody against Listeria monocytogenes, and Antibody 2 was goat anti-rabbit IgG against Listeria monocytogenes. Buy nano-Fe materials from the market, such as Beijing Deke Daojin Technology Co., Ltd. or Shanghai Chaowei Nano Materials Co., Ltd., 20nm, with a purity of 99.9%.

[0022] Nano-Fe coated with silicon dioxide: take 47.5g of sodium silicate, dissolve it in a beaker with deionized water, and adjust the pH value to 12-13 with hydrochloric acid. Take 5.0g of nano-Fe and add it into the beaker, and stir mechanically (with a glass rod) for 5min. The mixture was sonicated for 30 min and stirred in due course. Raise the temperature to 85°C, add hydrochloric acid dropwise to adjust the pH value to 6-7, and form a precipitate. While magnetically separating, wash the precipitate ...

example 2

[0029] Determination of food samples for the presence of harmful pathogenic bacteria Escherichia coli O157:H7.

[0030] 1. Preparation of nano-Fe immunoprobe: 1 anti-rabbit anti-IgG monoclonal antibody of O157:H7, and 2 anti-goat anti-rabbit IgG of O157:H7, which can be monoclonal antibody or polyclonal antibody. Fe nanoparticles can be purchased from the market (such as Chaowei Nanomaterials in Shanghai).

[0031] Nano-Fe coated with silicon dioxide: take 47.5g of sodium silicate, dissolve it in a beaker with deionized water, and adjust the pH value to 12-13 with hydrochloric acid. Take 5.0g of nano-Fe and add it into the beaker, and stir mechanically (with a glass rod) for 5min. The mixture was sonicated for 30 min and stirred in due course. Raise the temperature to 85°C, add hydrochloric acid dropwise to adjust the pH value to 6-7, and form a precipitate. While magnetically separating, wash the precipitate with deionized water for 3-4 times. Then, the precipitate was di...

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Abstract

The invention discloses an NMR rapid detection method for food-borne pathogenic bacteria based on the indirect enrichment of paramagnetic nanometer Fe probes, and belongs to the technical field of rapid detection of food-safe pathogenic bacteria. The present invention relies on the established nuclear magnetic resonance detection method that can be used for pathogenic bacteria in food liquid samples, utilizes 1 antibody to capture target bacteria, and utilizes the antibody of 1 antibody, namely, the paramagnetic nano-Fe probe prepared by 2 anti-coating to target bacteria Enrichment and separation are carried out, and the effect of the paramagnetic properties of nano-Fe on the relaxation time of the nuclear magnetic resonance attenuation signal is used to detect whether the sample contains target bacteria. The different specific correspondences are: paramagnetic nano-Fe probes show a linear relationship under certain conditions, that is, the larger the content of nano-Fe, the smaller the spin-lattice relaxation time or spin-spin relaxation time value of the sample , within a certain range can quantitatively detect the target bacteria. The method can be used for rapid detection of harmful pathogenic bacteria in food samples, and thus can be used as a rapid screening of a large number of samples to be tested.

Description

technical field [0001] The invention relates to a rapid detection method for pathogenic bacteria, in particular to an NMR rapid detection method for food-borne pathogenic bacteria based on the indirect enrichment of paramagnetic nanometer Fe probes. Background technique [0002] Basic principle: Monoclonal antibodies or antigen molecules are combined with enzyme molecules through covalent bonds. This combination will not change the immunological characteristics and biological activities of monoclonal antibodies, antigens and enzymes. Specific monoclonal antibodies will only interact with specificity. antigen binding. Fe material has ferromagnetism, and when the particle diameter is small to a certain extent, paramagnetic properties will appear. That is, there is no magnetism when there is no external magnetic field, but it shows certain magnetism when there is an external magnetic field, which can be used for magnetic separation. At the same time, the influence of paramagn...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N24/08G01N33/577
Inventor 张锦胜唐群赖卫华
Owner NANCHANG UNIV