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Quick detection method for food-borne pathogens based on immunomagnetic separation of Fe3O4 nano materials

A technology for food-borne pathogenic bacteria and immunomagnetic separation, which is used in analytical materials, measuring devices, instruments, etc.

Inactive Publication Date: 2013-05-22
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

[0019] Testing food samples for the presence of the harmful pathogen Listeria.

[0020]

[0021] 1. Preparation of immunomagnetic beads: coupling of magnetic beads and monoclonal antibodies. Use the carboxylated magnetic beads PM3-020 (180nm) commercialized by Aorun Company, or prepare Fe by yourself by microemulsion method 3 o 4 Nano-microspheres are modified into carboxylated magnetic beads or aminated magnetic beads by coating. The monoclonal antibody uses the commercial Listeria monoclonal antibody purchased by the biological reagent company, or prepares the monoclonal antibody by itself and separates and purifies it. Taking carboxylated magnetic beads as an example, coupling steps: ① Washing: 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 rinse with 2 mL of water after washing. Morpholineethanesulfonic acid (MES) 0.05M, pH = 6.0 resuspended; ② activation: Ultrasound to disperse the m...

example 2

[0027] Determination of food samples for the presence of the harmful pathogen Escherichia coli O157:H7.

[0028]

[0029] 1. Preparation of immunomagnetic beads: coupling of magnetic beads and monoclonal antibodies. Use carboxylated magnetic beads PM3-020 (180nm) commercialized by Aorun Company, or prepare Fe by yourself by microemulsion method 3 o 4 Nano-microspheres are modified into carboxylated magnetic beads or aminated magnetic beads by coating. The monoclonal antibody adopts the commercial E. coli O157:H7 monoclonal antibody, or prepares the monoclonal antibody by itself and separates and purifies it. Taking carboxylated magnetic beads as an example, coupling steps: ① Washing: Take 5 mg of carboxylated magnetic beads in a 5 mL centrifuge tube, wash twice with 2 mL of PB (0.02M, pH=6.0), wash with 2 mL of MES (0.05M , pH=6.0) resuspension; ②Activation: Ultrasonic to disperse the magnetic beads, then add 1mg EDC and 1mg NHSS (dissolved with 0.05M, pH=6.0 MES), activat...

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Abstract

The invention relates to a quick detection method for food-borne pathogens based on immunomagnetic separation of Fe3O4 nano materials, belonging to the technical field of quick detection of food safety pathogens. Depending on an established quick screening method which can be used for pathogens in a food liquid sample, the invention prepares immunomagnetic beads from Fe3O4 nano materials, enriches target strains in a targeted manner, and indirectly detects whether the target pathogens are contained in the sample by separating the nano magnetic beads capturing the target bacteria, converting into iron ions through nitration reaction and inspecting the iron ions. The amount of the iron ions and the target bacteria show a linear relationship under certain conditions, and the target bacteria can be quantitatively detected in a certain range. The method can be used for the quick detection of harmful pathogens in a food sample, and thus can be used for quickly screening a large number 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 immunomagnetic separation of nanomaterials. 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. Its main principle steps: 1. Utilize Fe 3 o 4 Magnetic beads, coated with a layer of polymer compounds, after modification, use an appropriate coupling agent to couple the monoclonal antibody of the target bacteria to be tested on the surface of the magnetic beads to form specific immunomagnetic beads and block redundant active sites . 2. Immobilize the specific monoclon...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
Inventor 张锦胜彭红赖卫华
Owner NANCHANG UNIV
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