Various mycotoxin quantifying detection protein chip and kit thereof
A mycotoxin and protein chip technology, which is applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of single function and slow efficiency, and achieve the effect of improving efficiency
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Embodiment 1
[0029] Preparation of a protein chip for quantitative detection of multiple mycotoxins
[0030] The main steps are: (1) Prepare the matrix nitrocellulose membrane; (2) Design the chip, determine the arrangement of the lattice and the spotting position; (3) Draw the antibody from the 384-well plate and spray it on the nitrocellulose (4) assemble a nitrocellulose membrane, and carry out sealing, drying, packaging, and preservation.
[0031] The above method for preparing a protein chip for quantitative detection of various mycotoxins specifically includes the following steps:
[0032] 1. Determination of mycotoxin detection antigen antibody
[0033] Determination of detection index antigen:
[0034] Select 7 kinds of mycotoxins that are more harmful and more widespread as antigens, extract and purify from raw materials. Take the extraction of aflatoxin as an example: the raw material containing aflatoxin will be extracted with a certain proportion of 40% methanol extract. Af...
Embodiment 2
[0042] Kit preparation
[0043] A kit for protein chips for quantitative detection of multiple mycotoxins, consisting of protein chips for quantitative detection of multiple mycotoxins, standard products, quality control products, sample diluents, enzyme-labeled working solutions, concentrated washing solutions, and detection solutions; each part It is bottled and fixed in the box with a paper template (as shown in the figure
[0044] 2), there is no limit to the placement method, and it is used in conjunction with the detection. Here's how the parts are prepared:
[0045] 1. The protein chip for the quantitative detection of multiple mycotoxins is a 6×8-well plate, with nitrocellulose membrane as the base, and the assembly method is shown in ZL032298900. After the completion of the appearance, see figure 1 .
[0046] 2. The standard product provides 6 bottles, each bottle is 0.25ml.
[0047]Prepare with phosphate buffer (pH 7.4) containing 3% BSA and 0.05% preservative (...
Embodiment 3
[0065] Application and detection of protein chips and kits
[0066] The application of the protein chip for the quantitative detection of various mycotoxins according to the present invention mainly adopts the antibody-antigen-enzyme-labeled secondary antibody method to detect the mycotoxins in the sample. The method is: solid samples such as grains, soybeans, etc. Crush, take 1g and put it into 1ml sample diluent, mix well, let the sample fully react with the sample diluent for half an hour, then absorb part and add it to the surface of the chip; for liquid samples such as juice, grain oil, etc., directly place 1ml in the 1ml of sample diluent, mix well, let the sample fully react with the sample diluent for half an hour, then absorb the part and add it to the surface of the chip. The mycotoxin in the sample, as an antigen-like component, binds to the corresponding specific antibody immobilized on the chip. After drying, add the enzyme-labeled secondary antibody to react, was...
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