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.
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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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