Method for detecting T-2 toxin and special reagent kit thereof

A kit, T-2 technology, applied in the direction of chemical instruments and methods, biochemical equipment and methods, measuring devices, etc., can solve the problems of lack of sensitivity, high price, complexity, etc., and achieve high sensitivity, low cost, and accuracy high 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

Some of these methods are too complex, some lack sensitivity, and some are expensive for routine toxin analysis

Method used

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  • Method for detecting T-2 toxin and special reagent kit thereof
  • Method for detecting T-2 toxin and special reagent kit thereof
  • Method for detecting T-2 toxin and special reagent kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1, preparation of kit components

[0040] Yield = the amount of main reactants consumed in conversion to products / the total consumption of main reactants × 100%.

[0041] 1. Antigen preparation

[0042] 1. Preparation of hapten (CMO-T-2)

[0043] The T-2 toxin hapten refers to a product (CMO-T-2) obtained by reacting T-2 toxin with O-(carboxymethyl) hydroxylamine hemihydrochloride. The specific steps of preparation are as follows:

[0044] ①Weigh 9.5mg of T-2 toxin and dissolve in 1ml of dry dichloromethane, called solution A; weigh 17mg of pyridinium chlorochromate (PCC) and dissolve in 2ml of dry dichloromethane, called solution B ;

[0045] ②At room temperature, under stirring, add liquid B to liquid A drop by drop, and stir for 66 hours;

[0046] ③The reaction mixture (black) was diluted with 5ml of dichloromethane, washed 4 times with 5ml of saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain 7.7mg of light yellow compou...

Embodiment 2

[0091] Example 2, the preparation, application and performance identification of the enzyme-linked immunosorbent assay kit for detecting vomitoxin

[0092] 1. The composition of the kit

[0093] 1. T-2 toxin standard solution

[0094] 6 bottles of T-2 toxin standard solution, the concentrations are 0ng / mL, 0.2ng / mL, 0.4ng / mL, 0.8ng / mL, 1.6ng / mL, 3.2ng / mL.

[0095] 2. Substrate chromogenic solution

[0096] 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).

[0097] 3. Termination solution

[0098] The stop solution is 2mol / L sulfuric acid.

[0099] 4. Concentrated washing liquid

[0100] 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.

[0101] 5. Sample concentrate

[0102] The sample concentrat...

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PUM

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Abstract

The invention discloses a method for detecting T-2 toxin and a special reagent kit thereof. The invention provides a hybridoma cell line 3E-5A-7H-8B whose preservation number is CGMCC No. 3395. The invention also provides a monoclonal antibody which is secreted by the hybridoma cell line 3E-5A-7H-8B whose preservation number is CGMCC No. 3395. The reagent kit provided by the invention comprises the monoclonal antibody. In the invention, the structure of the T-2 toxin is reconstructed, a space arm is added, 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 high specificity. The reagent kit of the invention has the characteristics of simpleness 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 T-2 toxin.

Description

technical field [0001] The invention relates to a method for detecting T-2 toxin and a special kit thereof. Background technique [0002] T-2 toxin (T-2toxin), belonging to trichothecene group A compounds, is one of the most toxic and highly polluting toxins in this class of compounds, mainly produced by Fusarium mould. T-2 toxin mainly contaminates grains such as wheat, corn, barley, oats and rye, and agricultural products such as malt, beer and bread. Most of the reports on the contamination of T-2 toxin focused on the reports on grain and feed. T-2 toxin is an inhibitor of protein synthesis in the body. After humans and animals eat sick grains by mistake, it can cause acute poisoning symptoms such as vomiting, diarrhea, and fever. In severe cases, it can damage hematopoietic tissues and cause casualties. In addition, T-2 toxin is also closely related to the high incidence of esophageal cancer, Keshan disease and Kaschin-Beck disease in some areas of my country. [0003...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/44G01N33/577
Inventor 吴小平江海洋王战辉史为民徐飞张兴祥王熙潘净如王进李娜
Owner 北京维德维康生物技术有限公司
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