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Method for detecting deoxynivalenol and special reagent kit thereof

The technology of vomitoxin and reagent kit is applied in the direction of chemical instruments and methods, biochemical equipment and methods, measuring devices, etc., which can solve problems such as human and animal health risks, and achieve low cost, high specificity, and high accuracy. Effect

Inactive Publication Date: 2011-06-01
北京维德维康生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the toxin poses a great danger to human and animal health

Method used

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  • Method for detecting deoxynivalenol and special reagent kit thereof
  • Method for detecting deoxynivalenol and special reagent kit thereof
  • Method for detecting deoxynivalenol and special reagent kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, preparation of kit components

[0044] Yield = the amount of main reactants consumed in conversion to products / the total consumption of main reactants × 100%.

[0045] 1. Antigen preparation

[0046] 1. Preparation of hapten (DON-succinic anhydride)

[0047] Dissolve vomitoxin in pyridine, add 1-butylboronic acid to seal the two hydroxyl groups on the 7th and 15th positions in the molecular structure of DON first, and then react the hydroxyl group on the 3rd position with succinic anhydride, so that on the DON molecule A spacer arm containing a carboxyl group was constructed to obtain the deoxynivalenol hapten. The specific steps are as follows: Weigh 2 mg of a standard sample of vomitoxin (DON), dissolve it in 200 uL of pyridine, add 20.8 mg of 1-butylboronic acid, stir the mixture at room temperature for 20 h, and then add 1.7 mol / L succinic anhydride while stirring The pyridine solution was 400uL, the reaction tube was sealed, the mixture was stirred...

Embodiment 2

[0097] Example 2, the preparation, application and performance identification of the enzyme-linked immunosorbent assay kit for detecting vomitoxin

[0098] 1. The composition of the kit

[0099] 1. Deoxynivalenol standard solution

[0100] Six bottles of deoxynivalenol standard solution, the concentrations were 0μg / L, 0.5μg / L, 2μg / L, 8μg / L, 32μg / L and 128μg / L.

[0101] 2. Substrate chromogenic solution

[0102] The substrate chromogenic solution is composed of A solution and B solution, the substrate chromogenic solution A is 2% carbamide peroxide solution, and the substrate chromogenic solution B is 1% tetramethylbenzidine solution (TMB).

[0103] 3. Termination solution

[0104] The stop solution is 2mol / L sulfuric acid.

[0105] 4. Concentrated washing liquid

[0106] The concentrated washing solution is a solution obtained by adding 1.0 mL Tween 20 and 0.5 g sodium azide to 100 mL pH7.4 0.01M phosphate buffer.

[0107] 5. Sample concentrate

[0108] The sample conce...

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Abstract

The invention discloses a method for detecting deoxynivalenol and a special reagent kit thereof. The invention provides a hybridoma cell line 8C-2H-3A-6F-2B whose preservation number is CGMCC No. 3396. The invention also provides a monoclonal antibody which is secreted by the hybridoma cell line 8C-2H-3A-6F-2B whose preservation number is CGMCC No. 3396. The reagent kit provided by the invention comprises the monoclonal antibody. In the invention, the structure of the deoxynivalenol is reconstructed, a space arm is added, the characteristic group of the deoxynivalenol is stressed, and artificial antigens are synthesized. The artificial antigens are used for immuning animals to obtain the hybridoma cell line, and the monoclonal antibody which is secreted by the hybridoma cell line has highspecificity. The reagent kit of the invention has the characteristics of simpleness and portability of operation, low manufacture cost, high specificity, high sensibility, high precision and the like, the reagent kit which can be used for field monitoring is suitable for screening a great number of samples and can play an important role in detecting the deoxynivalenol.

Description

technical field [0001] The invention relates to a method for detecting vomitoxin and a special kit thereof. Background technique [0002] The chemical name of vomitoxin is deoxynivalenol (Deoxynivalenol, DON), and its chemical structure is shown in formula (II): [0003] Formula (II). [0004] Deoxynivalenol belongs to trichothecene compounds and is mainly produced by Fusarium graminearum and Fusarium culmorum. It is also called vomitoxin because it can cause vomiting in animals. There are more than 150 kinds of trichothecene toxins, which are powerful immunosuppressants. The typical symptom caused by them is reduced feed intake, so this type of toxin is also called feed refusal toxin. DON is one of the most important toxins. [0005] DON is mostly distributed in grain seeds such as wheat, barley, and corn. It has strong cytotoxicity and has obvious toxic effects on prokaryotic cells, eukaryotic cells, plant cells, and tumor cells. In addition, vomitoxin can also act o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/44G01N33/577
Inventor 江海洋吴小平徐飞王战辉史为民王照鹏王奇昌潘净如
Owner 北京维德维康生物技术有限公司
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