ELISA kit for detecting trenbolone residue and detection method thereof

A technology of trenbolone and kits, applied in the field of enzyme-linked immunosorbent assay kits, which can solve the problems of cumbersome processing and measurement operations, high cost, and high cost of using enzyme-linked immunosorbent assay methods

Inactive Publication Date: 2006-05-03
贵州勤邦食品安全科学技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The gas chromatography (RPMC) analysis method sample pretreatment and determination operation is cumbersome and expensive, and the enzyme-linked immunosorbent assay method is expensive to use, the recovery method is complicated, and the sensitivity is not high. These defects limit its popularization and application.

Method used

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  • ELISA kit for detecting trenbolone residue and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Example 1 Detecting the preparation of the components of the ELISA kit for trenbolone

[0085] 1. Antigen Synthesis

[0086] a. Synthesis of Coating Progen

[0087] The trenbolone hapten was synthesized from trenbolone by the succinic anhydride method. Because there is a hydroxyl group in the molecular structure of trenbolone, a hapten with a similar structure can be designed by using the succinic anhydride method, and the prepared antigen highlights the original structure of the trenbolone molecule, making the prepared trenbolone antibody more specific. The coating agent is obtained by coupling hapten and bovine serum albumin (BSA) carrier protein by water-soluble carbodiimide method (EDC).

[0088] b. Synthesis of Immunogen

[0089] The trenbolone hapten was synthesized from trenbolone by the succinic anhydride method. Because there is a hydroxyl group in the molecular structure of trenbolone, a hapten with a similar structure can be designed by using the succinic...

Embodiment 2

[0104] Embodiment 2 detects the formation of the ELISA kit of trenbolone

[0105] An ELISA kit for detecting trenbolone was set up to include the following components:

[0106] (1) an enzyme-linked plate coated with trenbolone antigen;

[0107] (2) Goat anti-mouse anti-antibody labeled with horseradish peroxidase;

[0108] (3) Trenbolone mouse monoclonal antibody;

[0109] (4) 6 bottles of trenbolone standard solution, the concentrations are 0 μg / L, 0.05 μg / L, 0.15 μg / L, 0.45 μg / L, 1.35 μg / L, 4.05 μg / L;

[0110] (5) Chromogenic solution A liquid is hydrogen peroxide, and chromogenic liquid B liquid is o-phenylenediamine;

[0111] (6) The stop solution is 2mol / L sulfuric acid buffer;

[0112] (7) The concentrated washing solution is pH 7.4, containing 0.8% Tween 20 and 1‰ sodium azide (NaN 3 ) phosphate buffered saline solution with preservatives;

[0113] (8) The concentrated complex solution is a phosphate buffer containing 5‰N,N'-dimethylformamide (DMF).

Embodiment 3

[0114] Example 3 The formation of the ELISA kit for detection of trenbolone The ELISA kit for detection of trenbolone is formed so that it includes the following components:

[0115] (1) Enzyme-linked plate coated with goat anti-rabbit anti-antibody;

[0116] (2) Trenbolone antigen labeled with alkaline phosphatase;

[0117] (3) Trenbolone rabbit polyclonal antibody;

[0118] (4) 6 bottles of trenbolone standard solution, the concentrations are 0 μg / L, 0.05 μg / L, 0.15 μg / L, 0.45 μg / L, 1.35 μg / L, 4.05 μg / L;

[0119] (5) The substrate solution is p-nitrophosphate buffer;

[0120] (6) The stop solution is 2mol / L sodium hydroxide buffer solution;

[0121] (7) The concentrated washing liquid is pH 7.4, containing 1.2% Tween 20 and 1‰ sodium azide (NaN 3 ) phosphate buffered saline solution with preservatives;

[0122] (8) The concentrated complex solution is a phosphate buffer containing 5‰ N, N'-dimethylformamide (DMF).

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Abstract

The invention provides an enzyme immune agent box for detecting Kwan Bo Long of animal foodstuff which comprises: enzyme mark plate which coats Kwan Bo Long antigen or antibody, enzyme mark material, Kwan Bo Long peculiar antibody, Kwan Bo Long standard solution, base material color developing solution, compression cleaning liquid, ending solution and compression twin solution. The invention also discloses a method for applying the detecting method, which comprises: first doing sample front process, then using the agent box to detect, at last analyzing the detected result.

Description

technical field [0001] The invention relates to an ELISA kit and method for detecting trenbolone in animal-derived food. Background technique [0002] Trenbolone (Trenbolone, TRE) is a widely used steroid anabolic hormone, which once played an important role in the control and treatment of livestock and poultry diseases. However, because trenbolone has serious side effects and can cause diseases, developed countries such as Europe and the United States have successively banned or strictly prohibited its use. According to the No. 235 document of the Ministry of Agriculture of my country, the residue limit of trenbolone in food of animal origin shall not be detected. [0003] At present, the methods used for the detection of trenbolone residues mainly include gas chromatography (RPMC) analysis and enzyme-linked immunosorbent assay. The gas chromatography (RPMC) analysis method sample pretreatment and determination operation is cumbersome and expensive, and the enzyme-linked ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/52G01N33/535G01N33/543
Inventor 沈建忠何方洋万宇平冯才伟吴小平冯才茂汪善良李军赵正苗张照亮史为民张素霞丁双阳罗晓琴孙倩
Owner 贵州勤邦食品安全科学技术有限公司
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