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Chemiluminescence immune detection reagent kit for detecting Clenbuterol

A chemiluminescent immunoassay and detection kit technology, which is applied in the field of immunological detection, can solve the problems of prolonging the detection time and achieve the effects of increasing sensitivity, low detection cost and reducing incubation time

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 this method 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), and the incubation reaction is required after the addition of the two antibodies, 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 clenbuterol hapten: acidify clenbuterol and react with sodium nitrite in a low temperature environment without light at 4°C to generate an intermediate containing diazonium cations. The diazotized clenbuterol is used as a hapten for the subsequent synthesis of immunizing antigens and coating antigens.

[0051] Synthesis of bovine serum albumin-clenbuterol immunization antigen: Clenbuterol was coupled with bovine serum albumin (BSA) by diazotization method to obtain immunization antigen.

[0052] Synthesis of Human Serum Albumin-Clenbuterol Coated Antigen: Clenbuterol is coupled with human serum albumin (HSA) by diazotization method to obtain immunizing antigen.

[0053] Preparation of clenbuterol-peroxidase-labeled antibody: the above-mentioned immunized antigens were injected into Balb / C mice, and after multiple boosting immunizations, antibody serum was produced. The spleen cells of the immunized mi...

Embodiment 2

[0081] A chemiluminescent immunoassay kit for the detection of clenbuterol contains the following components:

[0082] (1) A 48-well opaque white microtiter plate (6 strips × 8 wells) was coated with Clenbuterol-mouse serum albumin cross-linked complex, and was vacuum-sealed with aluminum film.

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

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

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

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

[0087] (5) 50-fold concentrated buffer. Before use, it was diluted 50 times to become the working concentration buffer, and the diluted working concentration buffer was 0.05mol / LTris-HCl buffer, pH 6.9, containing 0.1% (v / v) Tween-20.

Embodiment 3

[0089] A chemiluminescent immunoassay kit for the detection of clenbuterol contains the following components:

[0090] (1) A 96-well opaque white microtiter plate (12 strips × 8 wells) was coated with Clenbuterol-chicken protein cross-linked complex, and was packaged with aluminum film vacuum-sealed.

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

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

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

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

[0095] (5) 10-fold concentrated buffer. Before use, it was diluted 10 times to become the working concentration buffer, and the diluted working concentration buffer was 0.05mol / L glycine-HCl buffer, pH 6.9, containing 0.02% (v / v) Tween-20.

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Abstract

The invention provides a chemiluminescent immunoassay kit for detecting clenbuterol, 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 clenbuterol-carrier protein cross-linked complex, a clenbuterol standard product, a clenbuterol-peroxidase-labeled antibody, luminescent substrate solution and concentrated buffer solution. The clenbuterol-carrier protein cross-linked complex is obtained by coupling the clenbuterol 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 in the detection of residual clenbuterol in animal urine, blood, tissues, visceral organs and other samples.

Description

technical field [0001] The invention relates to a chemiluminescence immunoassay kit for detecting clenbuterol, which is used for detecting the content or residue of clenbuterol in animal-derived food such as animal tissue, viscera, blood, urine and feed, feed raw materials quantity. It belongs to the field of immunological detection. Background technique [0002] Clenbuterol (CLB), commonly known as clenbuterol, is a beta-stimulant. Because it can improve the ratio of meat to fat in fatty animals and accelerate animal growth, it is widely added to animal feed and can remain in animals. However, this drug has serious side effects in humans. From light to lead to abnormal heartbeat and heart rhythm, in severe cases can lead to heart disease. At present, my country and many countries have banned its use as a feed additive. [0003] In terms of detection methods for clenbuterol, the most commonly used methods include high performance liquid chromatography (HPLC), gas chroma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/543G01N21/76
Inventor 陈峰徐晓婴卢庆鋆梁晓翠卢永红
Owner SHANGHAI JIAO TONG UNIV
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