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NMR detection method for food-borne pathogenic bacteria based on Fe3O4 nano-material

A technology of food-borne pathogens and detection methods, applied in the direction of analysis by nuclear magnetic resonance

Inactive Publication Date: 2013-07-17
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 the enrichment of targets using immunomagnetic beads, but none of them have used nuclear magnetic resonance for further detection

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0021] Test food samples for the presence of the harmful pathogenic bacteria Listeria.

[0022]

[0023] 1. Preparation of immunomagnetic beads: coupling of magnetic beads and monoclonal antibodies. Commercially available carboxylated magnetic beads from Aorun Company, PM3-020 (180nm) were coupled with monoclonal antibodies. The monoclonal antibody adopts the commercial Listeria monoclonal antibody purchased by the biological reagent company, or prepares the monoclonal monoclonal antibody by itself and separates and purifies it. Coupling steps: ① Cleaning: Take 5 mg of carboxylated magnetic beads in a 5 mL centrifuge tube, wash twice with 2 mL of phosphate buffer (PB) 0.02M, pH=6.0, and wash with 2 mL of morpholineethanesulfonic acid (MES) Resuspend at 0.05M, pH=6.0; ②Activation: Sonicate to disperse the magnetic beads, then add 1mg carbodiimide (EDC), 1mg N-hydroxysuccinimide (NHSS) (use 0.05M, pH=6.0 MES Dissolved), activated at 25°C for 2 hours, rotator 15r / min to mainta...

example 2

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

[0030]

[0031] 1. Preparation of immunomagnetic beads: coupling of magnetic beads and monoclonal antibodies. Commercially available carboxylated magnetic beads from Aorun Company, PM3-020 (180nm) were coupled with monoclonal antibodies. The monoclonal antibody adopts the commercial Escherichia coli O157:H7 monoclonal antibody, or prepares the monoclonal monoclonal antibody by itself and separates and purifies it. 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, was...

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Abstract

The invention provides a rapid detection method for food-borne pathogenic bacteria based on an NMR nanometer probe of Fe3O4, belonging to the field of rapid detection technology for pathogenic bacteria in food safety. According to the invention, based on an established nuclear magnetic resonance detection method applicable to detection of pathogenic bacteria in a liquid food sample, a Fe3O4 nano-material is used for preparing an immunomagnetic bead for specific enrichment of a target bacterium, and influence of paramagnetic and super-paramagnetic characteristics of Fe3O4 on nuclear magnetic resonance relaxation time is used for detecting whether the sample contains a target pathogenic bacterium or not. A different concrete corresponding relationship is that the super-paramagnetic immunomagnetic bead presents linear relation under certain conditions, i.e., the greater the content of the immunomagnetic bead is, the smaller a T2 / T1 value of the sample is, and quantitative detection of the target bacterium can be realized in a certain range. The method provided by the invention is applicable to rapid detection of harmful pathogenic bacteria in a food sample, so the method can be used for rapid screening of large quantities of to-be-detected samples.

Description

technical field [0001] The invention relates to a method for detecting pathogenic bacteria NMR, in particular to a method based on Fe 3 o 4 NMR detection method of foodborne pathogens using nanomaterials. Background technique [0002] Basic principle: The monoclonal antibody or antigen molecule is combined with the enzyme molecule through a covalent bond. This combination will not change the immunological characteristics and biological activities of the monoclonal antibody, antigen and enzyme. The specific monoclonal antibody will only interact with the specific Sexual antigen binding. Its main principle steps 1. Utilize Fe 3 o 4 The magnetic beads are coated with a layer of high molecular compound. After modification, the monoclonal antibody of the target bacteria to be tested is coupled to the surface of the magnetic beads with an appropriate coupling agent to form specific immunomagnetic beads. 2. Immobilize the specific monoclonal antibody of another part of the tar...

Claims

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Application Information

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IPC IPC(8): G01N24/08
Inventor 张锦胜唐群赖卫华
Owner NANCHANG UNIV
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