Bigeminy qualitative fungaltoxin colloidal gold immunochromatography test strip and preparation method
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A technology of immunochromatographic test strips and mycotoxins, which is applied in the field of immunochromatography, can solve the problems of long time consumption and complicated result determination methods, and achieves the effect of low cost.
Inactive Publication Date: 2017-05-17
ZHEJIANG FORESTRY UNIVERSITY
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[0004] The technical problem to be solved by the present invention is to provide a mycotoxin dual colloidal gold Immunochromatography test strip and preparation method thereof
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[0031] The preparation method of duplex colloidal gold qualitative immunochromatography test strip comprises the following steps:
[0032] 1. Preparation of colloidal gold antibody marker immobilization pad
[0033] According to the above method, the mycotoxin-specific monoclonal and rabbit polyclonal antibody colloidal gold-labeled complexes were obtained, and placed at 4°C for later use;
[0034]Fix the colloidal gold-labeled complex of mycotoxin-specific monoclonal and rabbit polyclonal antibodies on the antibody marker immobilization pad 2 by spraying, coating or soaking, and dry at 37°C for 2 hours for use;
[0035] 2. Spraying of detection line and quality control line: Take nitrocellulose membrane 3 and stick it on the strip-shaped polyvinyl chloride backboard as the base material, and spray the first detection line 5 on the nitrocellulose membrane 3 at the same time , quality control line 6, second detection line 7, wherein quality control line 6 is located between th...
Embodiment 1
[0037] Example 1: Preparation of ochratoxin A and zearalenone dual qualitative colloidal gold immunochromatographic test strips
[0038] Concrete preparation process is as follows:
[0039] 1. Preparation of colloidal gold-ochratoxin A monoclonal antibody marker, colloidal gold-zearalenone monoclonal antibody and colloidal gold-rabbit polyclonal antibody marker immobilization pad
[0040] Prepare and purify anti-ochratoxin A and zearalenone mouse monoclonal antibodies by conventional method, the concentrations are 5mg / mL and 3mg / mL respectively, purchase commercial rabbit polyclonal antibodies, the concentration is 10mg / mL, and reserve;
[0041] Preparation of antibody colloidal gold labeling complex: use 0.2M K 2 CO 3 To adjust the pH of the colloidal gold solution, take 50 mL of ochratoxin A monoclonal antibody diluted with 10 mM Tris-HCl solution (pH7.4) and quickly add 50 μg of ochratoxin A monoclonal antibody at room temperature for 30 min; %, after fully mixing for 30...
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Abstract
The invention relates to immunochromatography technologies, and aims at providing a bigeminy qualitative fungaltoxin colloidal gold immunochromatography test strip and a preparation method. The test strip comprises a black strip-like polyvinyl chloride backboard taken as a base material, wherein the surface of the test strip is successively provided with a sample pad, a colloidal gold antibody marker fixing pad, a nitrocellulose membrane and an adsorption pad along the length direction of the backboard, and the adjacent pad or the membrane are in overlap joint with each other; and the nitrocellulose membrane is provided with two detection lines and one quality control line which have the same width with that of the backboard and are separated from one another, wherein the quality control line is positioned between the two detection lines. According to the test strip provided by the invention, the colloidal gold, the cost of which is relatively low, is adopted as an antibody marker, a competition method principle is adopted to quickly detect fungaltoxin, the nitrocellulose membrane wraps the two detection lines simultaneously, the color depth can be compared by naked eyes, and qualitative detection can be carried out quickly on two kinds of fungaltoxin simultaneously. The chromatography test strip prepared has excellent sensitivity and stability, and is easy, convenient and fast to operate and low in cost, thereby being particularly suitable for rapid diagnosis on site.
Description
technical field [0001] The invention relates to immunochromatography technology, in particular to a dual qualitative immunochromatography test based on colloidal gold-labeled mycotoxin-specific monoclonal antibodies and visual judgment for simultaneous detection of two mycotoxins in food. Method of making paper strips. Background technique [0002] Food safety poisoning incidents that frequently break out at this stage, except for some caused by inorganic compounds such as potassium cyanide and arsenic, most of them come from biological toxins, microbial toxins are the most common, and most food poisoning is caused by bacterial toxins or mycotoxins caused. Among them, mycotoxin is a kind of small molecular secondary metabolites produced by fungi. Crops are susceptible to mycotoxin contamination during growth, harvesting, storage and transportation. More than 400 kinds of mycotoxins have been found, most of which are It has carcinogenic, teratogenic and mutagenic effects, a...
Claims
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