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ELISA kit for detecting ractopamine in animal derived food

A technology for ractopamine and animal detection, applied in the field of immunology detection, can solve the problems of residual confirmation analysis, unsuitable sample screening, high cost, etc., and achieve the effect of reducing the loss of enzyme activity

Active Publication Date: 2006-05-03
BEIJING WANGER BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the microbial detection method is economical and easy to operate, its sensitivity and specificity are limited when there are other microbial inhibitors in the sample; High sensitivity, but cumbersome sample pretreatment and determination operations, high cost, not suitable for screening a large number of samples, can be used as a confirmatory analysis of residues

Method used

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  • ELISA kit for detecting ractopamine in animal derived food

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] The preparation of embodiment 1 kit components

[0088] 1. Antigen synthesis

[0089] a. Dissolve 2 g of the ractopamine hapten in 20 ml of 0.5 M sodium hydroxide solution.

[0090] b. Dissolve 2 g of nitrogen hydroxysuccinamide in 8 ml of pure water and add it to the ractopamine hapten solution, and stir at room temperature for 2.5 hours.

[0091] c. Take 22 g of human serum albumin (HSA) or thyroid protein (BCG) carrier protein and dissolve it in 75 ml of pH9 carbonate buffer.

[0092] d. Add the carrier protein dropwise to the hapten solution and stir overnight at 4°C.

[0093] e. Dialyze the synthetic artificial antigen with 0.2M phosphate buffer for 7 days, and change the solution 3 to 4 times a day. Finally, the antigen is concentrated and stored or freeze-dried to obtain the ractopamine immunogen and the coating source.

[0094] Generally, the purity of the immunogen is required to be high. The higher the purity of the immunogen, the stronger the specificity ...

Embodiment 2

[0112] Embodiment 2 detects the formation of the ELISA kit of ractopamine

[0113] Construct the ELISA kit for detecting ractopamine to include the following components:

[0114] (1) A microtiter plate coated with ractopamine antigen;

[0115] (2) ractopamine mouse monoclonal antibody with a protein concentration of 0.5 μg / L;

[0116] (3) Goat anti-mouse anti-antibody labeled with horseradish peroxidase;

[0117] (4) 6 bottles of ractopamine standard solution, the concentrations were 0 μg / L, 0.5 μg / L, 1.5 μg / L, 4.5 μg / L, 13.5 μg / L, 40.5 μg / L;

[0118] (5) The substrate chromogenic solution A liquid is hydrogen peroxide, and the substrate chromogenic liquid B liquid is o-phenylenediamine;

[0119] (6) The concentrated washing solution is a phosphate buffer containing 0.8% Tween 20;

[0120] (7) The complex solution is a phosphate buffer containing 50% methanol, pH8.0, 0.1mol / L;

[0121] (8) The stop solution is 1mol / L sulfuric acid solution.

Embodiment 3

[0122] Embodiment 3 detects the formation of the ELISA kit of ractopamine

[0123] Construct the ELISA kit for detecting ractopamine to include the following components:

[0124] (1) A microtiter plate coated with goat anti-mouse anti-antibody;

[0125] (2) Ractopamine monoclonal antibody with a protein concentration of 5.0 μg / L;

[0126] (3) Ractopamine labeled with alkaline phosphatase;

[0127] (4) 6 bottles of ractopamine standard solution, the concentrations were 0 μg / L, 0.5 μg / L, 1.5 μg / L, 4.5 μg / L, 13.5 μg / L, 40.5 μg / L;

[0128] (5) The substrate chromogenic solution is p-nitrophosphate buffer;

[0129] (6) The concentrated washing solution is a phosphate buffer containing 1.2% Tween 20;

[0130] (7) The complex solution is a phosphate buffer containing 50% methanol, pH8.6, 0.2mol / L;

[0131] (8) The stop solution is 2mol / L sodium hydroxide solution.

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Abstract

The invention discloses an enzyme immune agent box for detecting lake drugs of animal foodstuff; it also provides a method for using the agent to detect the lake drugs residue. The agent box comprises: enzyme mark plate which coats lake drugs antigen or antibody, lake drugs mouse monoclonal antibody or polyclonal antibody working solution, enzyme mark antibody or enzyme mark lake drugs antigen, lake drugs 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 the field of immunological detection, and specifically provides an enzyme-linked immunoimmunoassay kit for detecting ractopamine in animal-derived foods and a detection method thereof. Background technique [0002] Ractopamine is a phenylethanolamine drug. Used in animal husbandry and fishery, it is a "nutritional redistribution agent", which belongs to β-stimulants. As a substitute for "clenbuterol", ractopamine has good effect, high economic return, and is widely used; used in medicine, it is a A cardiotonic drug that can be used to treat obesity and muscular dystrophy. However, when the cumulative dose of human intake exceeds a certain value, or when ractopamine remains high in the internal organs, a poisoning reaction occurs. The symptoms are increased skeletal muscle contraction, destruction of the fusion between fast-twitch muscle fibers and slow-twitch muscle fibers, and muscle tremors. The muscles of the limbs and fac...

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张素霞史为民丁双阳罗晓琴孙倩
Owner BEIJING WANGER BIOTECH
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