Chemiluminescence detection kit of fumonisin and preparation method thereof

[0026] Example 1: Construction and preparation of kit 1

[0027] 1. Set up kit 1

[0028] Build a chemiluminescence detection kit for fumonisins, which contains the following components:

[0029] Fumonisin antibody labeled with acridinium ester;

[0030] Fumonisin antigen coupled with carboxyl magnetic beads;

[0031] Chemiluminescence pre-excitation solution A and chemiluminescence excitation solution B;

[0032] Fumonisin series calibrator solutions, the concentrations are: 0 ng / mL, 0.01 ng / mL, 0.1 ng / mL, 1 ng / mL, 10ng / mL, 200 ng / mL;

[0033] The cleaning solution is specifically a Tris-HCl solution with a pH of 7.2 and a concentration of 25 mmol / L, which contains NaCl with a concentration of 0.15 mol / L and 0.05% Tween-20.

[0034] 2. Preparation of magnetic particle suspension coupled with fumonisin antigen

[0035] (1) Take 1 mg of carboxyl magnetic particles in a 0.5 mL centrifuge tube, add 200 μL of 0.1 mol / L MES (pH 5.0) buffer, vortex to mix, place on a magnetic stand

[0050] Example 2: Construction and preparation of kit 2

[0051] 1. Construction of Kit 2

[0052] Build a chemiluminescence detection kit for fumonisins, which contains the following components:

[0053] Fumonisin antigen labeled with acridinium ester;

[0054] Carboxylic magnetic beads coupled with fumonisin antibody;

[0055] Chemiluminescence pre-excitation solution A and chemiluminescence excitation solution B;

[0056] Fumonisin series calibrator solutions, the concentrations are: 0 ng / mL, 0.01 ng / mL, 0.1 ng / mL, 1 ng / mL, 10ng / mL, 200 ng / mL;

[0057] The cleaning solution is specifically a Tris-HCl solution with a pH of 7.2 and a concentration of 25 mmol / L, which contains NaCl with a concentration of 0.15 mol / L and 0.05% Tween-20.

[0058] 2. Preparation of magnetic particle suspension conjugated with fumonisin antibody

[0059] (1) Take 1 mg of carboxyl magnetic particles in a 0.5 mL centrifuge tube, add 200 μL of 0.1 mol / L MES (pH 6.0) buffer, vortex to mix, place on a

[0074] Example 3: Detection of fumonisins in actual samples

[0075] The operating procedure of the fumonisin quantitative detection kit of the present invention is as follows:

[0076] 1. Sample preparation

[0077] (1) Corn flour sample processing

[0078] Take 1 g of the homogenized sample in a 50 mL centrifuge tube, add 10 mL of 70% methanol-PBS solution, shake vigorously, sonicate for 20 min, let stand for 15 min, centrifuge at 5000 r / min for 8 min, and take 50 μL of the supernatant Put it in a clean glass test tube, add 450μL of pH 8.0 0.1 mol / L phosphate buffer, and mix well to obtain the sample solution to be tested.

[0079] (2) Liquor sample processing

[0080] Take 200 μL of the sample in a centrifuge tube, add 980 μL of pH 8.0 phosphate buffer with a concentration of 0.1 mol / L, and mix well to obtain the sample solution to be tested.

[0081] 2. Testing of the kit

[0082] (1) Add 100 μL of the sample to be tested, 150 μL of the suspension of coupled magnetic particles, and