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A rapid detection method for foodborne pathogens based on indirect enrichment immunomagnetic separation of Fe3O4 nanoparticles

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: 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, 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

[0018] Testing food samples to determine whether they contain the harmful pathogenic bacterium Listeria.

[0019] 1.1.2 Preparation of anti-immune magnetic beads: Antibody 1 uses rabbit anti-IgG monoclonal antibody against Listeria, and antibody 2 uses goat anti-rabbit IgG against 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 5 mg carboxylated magnetic beads in a 5 mL centrifuge tube, wash twice with 2 mL LPB (0.02M, pH=6.0), and resuspend with 2 mL MES (0.05 M, pH=6.0) after washing; ② Activate : Ultrasound to disperse the magnetic beads, then add 1mgEDC, 1mgNHSS (dissolved with 0.05M, pH=6.0MES), activate at 25°C for 2h, and keep the suspension at 15r / min with a rotator; ③Coupling: Magnetic separation, suck off the su...

example 2

[0024] Determination of food samples for the presence of the harmful pathogen Escherichia coli O157:H7.

[0025] 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 5 mg carboxylated magnetic beads in a 5 mL centrifuge tube, wash twice with 2 mL LPB (0.02M, pH=6.0), and resuspend with 2 mL MES (0.05 M, pH=6.0) after washing; ② Activate : Ultrasound to disperse the magnetic beads, then add 1mgEDC, 1mgNHSS (dissolved with 0.05M, pH=6.0MES), activate at 25°C for 2h, and keep the suspension at 15r / min with a rotator; ③Coupling: Magnetic separation, suck off the supernatant , wash once ...

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PUM

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Abstract

The invention discloses a method for rapid detection of food-borne pathogenic bacteria based on indirect enrichment immunomagnetic separation of Fe3O4 nanoparticles, which belongs to the technical field of rapid detection of food-safe pathogenic bacteria. The present invention relies on the established detection method for pathogenic bacteria in food liquid samples, uses 1 anti-specific binding target bacteria, and uses Fe3O4 nanoparticle materials to prepare 2 anti-immune magnetic beads to specifically enrich 1 anti-labeled targets For bacteria strains, the nano-magnetic beads that capture the target bacteria are separated, and then converted into iron ions by nitrification reaction, and the iron ions are tested to indirectly detect whether the sample contains the target pathogenic bacteria. Under certain conditions, the amount of iron ions has a linear relationship with the target bacteria, and the target bacteria can be quantitatively detected within a certain range. 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 present invention involves a fast detection party of pathogenic bacteria, especially a FE -based 3 O 4 Nano particles indirectly enrich the rapid detection method of food -based pathogenic bacteria for immune magnetic separation. Background technique [0002] Basic principles: monoclonal antibodies or antigen molecules are combined with covalent bonds through covalent bonds. This combination will not change the immunological characteristics and biological activity of monoclonal antibodies, antigens and enzymes. Specific monoclonal antibodies will only be with specificity.The antigen binding.The main principle steps: 1. Add the first antibody of the target bacteria to the sample to the test. 3 O 4 Nano particles, by modifying the surface functionalization of the antibody, the 1 antibody, the 2 antibody, is coupled on the surface of the magnetic bead to form a specific immune magnetic bead and closed the excess activity site.2. Fix the specificity of the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/02
Inventor 张锦胜唐群赖卫华
Owner NANCHANG UNIV