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Chemiluminescence immune detection reagent kit for detecting ractopamine

A technique of chemiluminescence immunity and ractopamine, applied in the field of immunological detection, can solve problems such as prolonging detection time, achieve the effects of increasing sensitivity, simple instrument, and ensuring long-term effectiveness

Inactive Publication Date: 2009-04-22
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of the method used by Xu Chuanlai et al. is that the primary antibody and the secondary antibody need to be used at the same time (the secondary antibody is an enzyme-labeled antibody). After the addition of the two antibodies, incubation is required, which prolongs the detection time.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] 1) Preparation of each component of the kit

[0050] Preparation of ractopamine hapten: acidify ractopamine, react with sodium nitrite in a low-temperature environment without light at 4°C, and generate intermediates containing diazonium cations. The diazotized ractopamine was used as a hapten for subsequent synthesis of immune antigens and coating antigens.

[0051] Synthesis of bovine serum albumin-ractopamine immune antigen: the immune antigen was obtained by coupling ractopamine and bovine serum albumin (BSA) by diazotization method.

[0052] Synthesis of human serum albumin-ractopamine-coated antigen: the immunization antigen was obtained by coupling ractopamine and human serum albumin (HSA) by a diazotization method.

[0053] Preparation of ractopamine-peroxidase-labeled antibody: inject the above-mentioned immunization antigen into Balb / C mice, and make it produce antibody serum after several booster immunizations. Take out the splenocytes of the immunized mice...

Embodiment 2

[0081] The chemiluminescent immunoassay kit for detecting ractopamine includes the following components:

[0082] (1) 48-well opaque white ELISA plates (6 strips x 8 wells) are coated with ractopamine-mouse serum albumin cross-linked complex, and are vacuum-sealed with aluminum film.

[0083] (2) 5 bottles of ractopamine standard solution, the concentrations are respectively:

[0084] 0μg / L, 0.04μg / L, 0.36μg / L, 3.24μg / L, 29.16μg / L

[0085] (3) Ractopamine-horseradish peroxidase labeled antibody solution.

[0086] (4) Luminescence substrate A solution (luminol and enhancer), and luminescence substrate B solution (urea hydrogen peroxide).

[0087] (5) 50 times concentrated buffer. Diluted 50 times before use to become a working concentration buffer, the diluted working concentration buffer is 0.05mol / L Tris-HCl buffer, pH6.9, containing 0.1% (v / v) Tween-20.

Embodiment 3

[0089] The chemiluminescent immunoassay kit for detecting ractopamine includes the following components:

[0090] (1) 96-well opaque white ELISA plate (12 strips x 8 wells) coated with ractopamine-egg albumin cross-linked complex, and vacuum-sealed with aluminum film.

[0091] (2) 6 bottles of ractopamine standard solution, the concentrations are respectively:

[0092] 0μg / L, 0.05μg / L, 0.25μg / L, 1.25μg / L, 6.25μg / L, 31.25μg / L.

[0093] (3) Ractopamine-horseradish peroxidase labeled antibody solution.

[0094] (4) Luminescence substrate A solution (luminol and enhancer), and luminescence substrate B solution (urea hydrogen peroxide).

[0095] (5) 10 times concentrated buffer. Diluted 10 times before use to become a working concentration buffer, the diluted working concentration buffer is 0.05mol / L glycine-HCl buffer, pH6.9, containing 0.02% (v / v) Tween-20.

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Abstract

The invention provides a chemiluminescent immunoassay kit for detecting ractopamine, and the kit belongs to the field of immunoassay. The kit is composed of a non-transparent white enzyme-labeled plate which is coated by a ractopamine-carrier protein cross-linked complex, a ractopamine standard product, a ractopamine-peroxidase-labeled antibody, luminescent substrate solution and concentrated buffer solution. The ractopamine-carrier protein cross-linked complex is obtained by coupling the ractopamine and a carrier protein through the mixed-anhydride method or the direct protein activation method, and the concentrated buffer solution contains Tween-20 buffer solution. The kit has the advantages of rapidness, simpleness, high sensitivity, low detection cost, good repeatability, high throughput and the like, and the kit can be used for detecting the residual ractopamine in animal urine, blood, tissues, visceral organs and other samples.

Description

technical field [0001] The invention relates to a chemiluminescent immunoassay kit for detecting ractopamine, which is used for detecting the content or residual amount of ractopamine in animal-derived foods such as animal tissue, viscera, blood, urine, feed, and feed raw materials. It belongs to the field of immunological detection. Background technique [0002] Ractopamine is a β-agonist. β-agonist is a kind of nutrient redistribution agent, which is a kind of phenylethanolamine derivatives similar in structure and function to adrenaline and norepinephrine. It can improve the ratio of meat to fat in fatty animals and reduce ketone bodies Fat content, increase lean meat rate, and can accelerate animal growth, so it is added to animal feed. Common β2-agonists include clenbuterol, ractopamine, and salbutamol. With the increasing supervision of clenbuterol (commonly known as "clenbuterol") in my country, the use of clenbuterol has gradually decreased, and the use of other β...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/543G01N21/76
Inventor 陈峰卢庆鋆徐晓婴梁晓翠卢永红
Owner SHANGHAI JIAO TONG UNIV
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