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A kind of vomitoxin quantitative detection kit and its application method

A quantitative detection and vomitoxin technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve problems such as difficulty in detecting vomitoxins, and achieve the effect of simple operation

Active Publication Date: 2016-05-25
ZHANGJIAGANG EENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is still a lack of kits that can quickly, efficiently, and highly sensitively detect DON, so it is still difficult to quickly and efficiently detect DON

Method used

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  • A kind of vomitoxin quantitative detection kit and its application method
  • A kind of vomitoxin quantitative detection kit and its application method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] The preparation method of the vomitoxin quantitative detection kit of the present invention, the specific steps are as follows:

[0059] 1. Preparation of magnetic separation reagent:

[0060] Select 100 mg of magnetic separation particles (0.1 μm in diameter) with superparamagnetic properties and carboxyl (COOH-) active groups on the surface, and use 0.1 mol / L 2-(N-morpholino)ethanesulfonic acid MES, pH 4.5 solution 10 mL After washing 3 times, the magnetic particles were resuspended in 1 mL of this solution. Then add 50 μL of 10 mg / ml EDC to activate, mix at 25°C for 2 hours, add 2 mg of anti-FITC antibody (goat antibody), and mix at 37°C for 2 hours. Then add an equal volume of 0.01mol / LPBS5%BSA (pH7.4) buffer and block at 37° for 1 hour; finally wash the magnetic beads 3 times with 10mL of 0.01mol / LPBS (pH7.4) buffer containing 0.5% BSA solution, And use this solution to make magnetic separation reagent solution.

[0061] The preparation method of described MES s...

Embodiment 2

[0084] 1. Preparation of magnetic separation reagent:

[0085] Select 100 mg of magnetic separation particles (0.2 μm in diameter) with superparamagnetic properties and carboxyl (COOH-) active groups on the surface, wash with 0.1 mol / LMES, pH4.5 solution 10 mL for 3 times, and resuspend the magnetic particles with 1 mL of this solution . After that, add 75ul of 10mg / ml EDC for activation, mix at 25°C for 2 hours, add 2mg of anti-FITC antibody (mouse anti), and mix at 37°C for 2 hours. Then add an equal volume of 0.01mol / LPBS5%BSA (pH7.4) buffer and block at 37° for 1 hour; finally wash the magnetic beads 3 times with 10mL of 0.01mol / LPBS (pH7.4) buffer containing 0.5% BSA solution, And use this solution to make magnetic separation reagent solution.

[0086] The preparation method of described MES solution is:

[0087] Weigh MES19.52g, Tris1.56g, NaCl4.24g in a 1L beaker, measure 950mL of purified water to dissolve, adjust the pH to 4.5; add purified water to make up to 1L, ...

Embodiment 3

[0109] 1. Preparation of magnetic separation reagent:

[0110] Select 100 mg of magnetic separation particles (diameter 0.1um) with superparamagnetic properties and carboxyl (COOH-) active groups on the surface, wash 3 times with 10 mL of 0.1 mol / LMES, pH4.5 solution, and resuspend the magnetic particles with 1 mL of this solution . Then add 100ul of 10mg / ml EDC to activate, mix for 2 hours at 25°C, add 2mg of anti-FITC antibody (rabbit anti), and mix for 2 hours at 37°C. Then add an equal volume of 0.01mol / LPBS5%BSA (pH7.4) buffer and block at 37° for 1 hour; finally wash the magnetic beads 3 times with 10mL of 0.01mol / LPBS (pH7.4) buffer containing 0.5% BSA solution, And use this solution to make magnetic separation reagent solution.

[0111] The preparation method of described MES solution is:

[0112] Weigh MES19.52g, Tris1.56g, NaCl4.24g in a 1L beaker, measure 950mL of purified water to dissolve, adjust the pH to 4.5; add purified water to make up to 1L, use a pore si...

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Abstract

The invention discloses a vomitoxin quantitative determination kit and a use method thereof. The vomitoxin quantitative determination kit comprises a magnetic separation reagent 1, a resist 2, an enzyme-labelled antigen agent 3, a cleaning solution 4, a calibrator and a quality control 5, a substrate solution 6 and a stop solution 7, wherein the magnetic separation reagent is a magnetic particle suspension combined with an FITC(fluorescein isothiocyanate)-DON antibody; the resist is an FITC-labelled anti-DON antibody solution, and the enzyme-labelled antigen agent is a DON solution coupled with a biological enzyme. The vomitoxin quantitative determination kit have the remarkable advantages of simple, quick, sensitive and accurate operation, and suitability for detecting vomitoxin samples in food or feed on a large scale daily.

Description

technical field [0001] The invention relates to the field of biological monitoring, and belongs to the field of biotechnology, in particular to a quantitative detection kit of vomitoxin and its application method. Background technique [0002] Vomitoxin (Vomitoxin), also known as deoxynivalenol (Deoxynivalenol, DON), the chemical name is 3α, 7α, 1,5-trihydroxyfusarium-9-en-8-one, which is a single-ended Sporane family of compounds. , DON has high cytotoxicity and immunosuppressive properties, therefore, it poses a threat to human and animal health. [0003] The magnetic particle separation enzyme-linked immunoassay technology, which was promoted in the 1990s, is a new type of enzyme-linked immunoassay method established by combining immunomagnetic particle separation technology and enzyme-linked immunoassay technology. The ELISA reaction of magnetic particles is carried out in a liquid phase, and the surface area of ​​the immune reaction is greatly improved compared with t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/533
CPCG01N33/535G01N33/54333
Inventor 王毅谦邵景东吴福平郭旸申进玲
Owner ZHANGJIAGANG EENTRY EXIT INSPECTION & QUARANTINE BUREAU