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Food-borne pathogenic bacteria quick detection method based on Gamma-Fe2O3@Au nano particle indirect enrichment and immunomagnetic separation

A food-borne pathogenic bacteria, immunomagnetic separation technology, applied in the direction of measuring devices, analysis materials, instruments, etc.

Inactive Publication Date: 2013-07-03
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, but there are still many reports on the enrichment of pathogenic bacteria and target objects using immunomagnetic beads, but there is no further detection using the method of detecting iron ions

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0022] Testing food samples to determine whether they contain the harmful pathogenic bacterium Listeria.

[0023] 1. Preparation of 2 anti-immune magnetic beads: 1 anti-listeria rabbit anti-IgG monoclonal antibody, 2 anti-goat anti-rabbit IgG Listeria monoclonal antibody, which can be monoclonal antibody or polyclonal antibody. gamma -Fe 2 o 3 Nanoparticles can be prepared by chemical co-precipitation or by other methods. If 1mL of diluted gamma -Fe 2 o 3 Mix with the same volume of 0.1 mol / L sodium citrate, dilute to 20mL, add excess 80 mmol / L NH 2 Add 0.1% (by weight) HAuCl dropwise to the OH·HCl solution 4 Solution 2mL, reacted for 1 h, washed and separated to obtain gamma -Fe 2 o 3 Au.

[0024] immunity gamma -Fe 2 o 3 Au preparation: gamma -Fe 2 o 3 After Au concentration, excess goat anti-rabbit IgG antibody was added, after incubation and washing, excess BSA was added to seal the surface active vacancies, washed, resuspended, and stored at 4°C until use...

example 2

[0030] Determination of food samples for the presence of the harmful pathogen Escherichia coli O157:H7.

[0031] 1.2 Preparation of anti-immune magnetic beads: Antibody 1 uses O157:H7 rabbit anti-IgG monoclonal antibody, and Antibody 2 is goat anti-rabbit IgG O157:H7, which can be monoclonal antibody or polyclonal antibody. gamma -Fe 2 o 3 Nanoparticles can be prepared by chemical co-precipitation or by other methods. If 1mL of diluted gamma -Fe 2 o 3 Mix with the same volume of 0.1 mol / L sodium citrate, dilute to 20mL, add excess 80 mmol / L NH 2 Add 0.1% (by weight) HAuCl dropwise to the OH·HCl solution 4 Solution 2mL, reacted for 1 h, washed and separated to obtain gamma -Fe 2 o 3 Au.

[0032] immunity gamma -Fe 2 o 3 Au preparation: gamma -Fe 2 o 3 After Au concentration, add excess O157:H7 goat anti-rabbit IgG antibody, incubate and wash, add excess BSA to seal surface active vacancies, wash, resuspend, and store at 4°C until use. The following method can ...

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Abstract

A food-borne pathogenic bacteria quick detection method based on Gamma-Fe2O3@Au nano particle indirect enrichment and immunomagnetic separation, and belongs to the technical field of quick pathogen detection for food security. On dependence of a method for detecting pathogen in a food liquid sample, the specificity of an antibody I is combined with the target bacteria, the Gamma-Fe2O3@Au composite nano particle is utilized to prepare immuomagnetic beads of an antibody II to enrich target strains marked by the antibody I, the nano magnetic beads of the target bacteria are captured through separation, and then subjected to nitration to be converted into ferrous ions, and the ferrous ions are detected to indirectly detect whether the sample contains target pathogen. Under a certain condition, the quantity of the ferrous ions and the target bacteria show linear relation, so that the target bacteria can be quantificationally detected within a certain range. The food-borne pathogenic bacteria quick detection method can be used for quick detection of harmful pathogen in the food sample, and can be used for quick screening of large quantities of samples to be detected.

Description

technical field [0001] The invention relates to a rapid detection method for pathogenic bacteria, in particular to a method based on γ-Fe 2 o 3 Rapid detection of foodborne pathogenic bacteria by indirect enrichment of Au nanoparticles by immunomagnetic separation. 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. The main principle steps: 1. Add the first antibody of the target bacterium to the sample to be tested. If the target bacterium exists, it will combine with the first antibody to form an antibody complex; 2. Use the seed growth method to prepare a magnetic gamma -Fe 2 o 3 For Au core-shell nanoparticles, the surface is functionalized by modi...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/577
Inventor 张锦胜唐群赖卫华
Owner NANCHANG UNIV
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