Preparation of ELISA kit for detecting deoxynivalenol

A technology of vomitoxin and kit, applied in the field of detection, can solve problems such as backward development, and achieve the effect of simple operation and sensitive detection.

Inactive Publication Date: 2014-05-21
JIANGSU WISE SCI & TECH DEV
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  • Abstract
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Problems solved by technology

At present, most of the immunological detection kits for vomitoxin are imported, while the development of domestic kits for vomitoxin is still relatively backward

Method used

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  • Preparation of ELISA kit for detecting deoxynivalenol

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Embodiment Construction

[0034] ELISA kit for detecting vomitoxin, which includes enzyme plate, vomitoxin standard, vomitoxin antibody working solution, enzyme-labeled secondary antibody working solution, substrate solution A, substrate solution B, stop solution and concentrated washing solution.

[0035] The preparation of the ELISA kit for detecting vomitoxin in the present invention is described in detail below,

[0036] Preparation of DON protein conjugates:

[0037]Dissolve 20 mg of DON in 200 μL of pyridine, add it to a 2 ml reaction bottle, and add 68 mg of 1-butylboronic acid to the reaction bottle. The mixture was stirred overnight at room temperature, yielding 7,15-oxo-(butylboron)-deoxynivalenol. Then add 32 mg of succinic anhydride therein, and pass through oxygen-free nitrogen. The reaction bottle was sealed, and the mixture was stirred in a boiling water bath for 90 min to produce 3-oxo-succinyl-7,15-oxo-(butylboron)-deoxynivalenol. After drying pyridine at room temperature, a small a...

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Abstract

The invention relates to preparation of an ELISA kit for detecting deoxynivalenol. The kit with sensitive detection, accuracy, fastness, simple operation and strong specificity is suitable for determination of a large number of samples. The kit includes an ELISA plate coated with deoxynivalenol antigens, a deoxynivalenol standard sample, a deoxynivalenol antibody working solution, an enzyme-labeled?second antibody working solution, a substrate solution A, a substrate solution B, a stop solution, a concentrating sample weak solution and a washing concentrate. The principle of the deoxynivalenol detection kit is solid phase indirect competitive enzyme-linked immunosorbent assay. An extracted sample, the enzyme-labeled?standard second antibody working solution and the antibody working solution are added to corresponding enzyme-labeled holes; after incubation for a period of time, the substrate solution A and the substrate solution B are added into a washing plate; under the action of the enzyme, blue color occurs in the holes; and the stop solution adding is added in, and the color changes from blue to yellow. The degree of color is in inverse proportion relationship with the content of deoxynivalenol in the standard substance or sample. The method can be directly used for detecting deoxynivalenol in cereals and cereal products.

Description

technical field [0001] The invention belongs to the technical field of detection, in particular to an ELISA kit for quantitative detection of vomitoxin residues in grains and grain products. Background technique [0002] Mycotoxins are secondary metabolites produced by fungi. There are currently more than 300 known mycotoxins with different chemical structures, among which more than 30 metabolites produced by fungal strains are pathogenic to both animals and humans. According to FAO estimates, about 25% of the world's food crops are polluted by mycotoxins every year, causing economic losses of hundreds of billions of dollars each year. Today, as humans are increasingly concerned about the quality and safety of food hygiene, the contamination of mycotoxins has become a problem that cannot be ignored. The common and relatively harmful mycotoxins in my country's agricultural products and feed mainly include aflatoxin, deoxynivalenol, deoxynivalenol, zearalenone and ochratoxin ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/531
CPCG01N33/56911
Inventor 杜道林曾昆
Owner JIANGSU WISE SCI & TECH DEV
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