Food-borne pathogenic bacteria quick detection method based on Fe3O4 nano particle indirect enrichment and immunomagnetic separation

A technology for food-borne pathogenic bacteria and immunomagnetic separation, which is applied in the direction of testing food, material inspection products, 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 path

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0021] Testing food samples to determine whether they contain the harmful pathogenic bacterium Listeria.

[0022] 1. 1. 2 Preparation of anti-immune magnetic beads: 1 anti-rabbit anti-IgG monoclonal antibody of Listeria, 2 anti-goat anti-rabbit IgG of Listeria, which can be monoclonal antibody or polyclonal antibody. 2 Preparation of anti-immune magnetic beads: coupling of magnetic beads and monoclonal antibodies. The commercial carboxylated magnetic beads of Aorun Company, PM3-020 (180nm) and 2 anti-coupling were used. Coupling steps: ① Cleaning: Take 5mg of carboxylated magnetic beads in a 5mL centrifuge tube, wash twice with 2mL PB (0.02M, pH=6.0), and resuspend with 2mL MES (0.05M, pH=6.0) after washing; ②Activation: Ultrasonic to disperse the magnetic beads, then add 1mg EDC and 1mg NHSS (dissolved with 0.05M, pH=6.0 MES) respectively, activate at 25°C for 2h, and keep the suspension at 15r / min on the rotator; ③Coupling: Magnetic separation, Aspirate the supernatant, wa...

example 2

[0028] Determination of food samples for the presence of the harmful pathogen Escherichia coli O157:H7.

[0029] 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. 2 Preparation of anti-immune magnetic beads: coupling of magnetic beads and monoclonal antibodies. The commercial carboxylated magnetic beads of Aorun Company, PM3-020 (180nm) and 2 anti-coupling were used. Coupling steps: ① Cleaning: Take 5mg of carboxylated magnetic beads in a 5mL centrifuge tube, wash twice with 2mL PB (0.02M, pH=6.0), and resuspend with 2mL MES (0.05M, pH=6.0) after washing; ②Activation: Ultrasonic to disperse the magnetic beads, then add 1mg EDC and 1mg NHSS (dissolved with 0.05M, pH=6.0 MES) respectively, activate at 25°C for 2h, and keep the suspension at 15r / min on the rotator; ③Coupling: Magnetic separation, Aspirate the supernat...

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PUM

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Abstract

A food-borne pathogenic bacteria quick detection method based on Fe3O4 nano particle indirect enrichment and immunomagnetic separation 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 Fe3O4 nano particle material 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 Fe 3 o 4 Rapid detection method of foodborne pathogens by indirect enrichment of nanoparticles and 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 magnetic Fe 3 o 4 For nanoparticles, the surface is functionalized by modifying the antibody, and the antibody of the 1st antibody, tha...

Claims

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Application Information

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IPC IPC(8): G01N33/02
Inventor 张锦胜唐群赖卫华
Owner NANCHANG UNIV
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