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ELISA kit for detecting stilbestrol residue and detection method thereof

A kit, the technology of diethylstilbestrol, is applied in the direction of biological testing, measuring devices, material inspection products, etc., which can solve the problems of unsuitability for large-scale sample screening, restrictions on promotion and use, and high cost

Inactive 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

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; although the gas chromatography analysis method has high sensitivity, the sample pretreatment and determination operations are cumbersome and costly. It is suitable for the screening of a large number of samples, but it can only be used as a confirmatory analysis of residues, and its promotion and use are limited

Method used

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  • ELISA kit for detecting stilbestrol residue and detection method thereof

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Comparison scheme
Effect test

Embodiment 1

[0073] Example 1 Component preparation of the ELISA kit for detecting diethylstilbestrol

[0074] 1. Antigen Synthesis

[0075] a. Synthesis of Coating Progen

[0076] Diethylstilbestrol and sodium chloroacetate are synthesized into a hapten through a halogen substitution reaction, and a spacer arm containing 2 carbon chains is connected to diethylstilbestrol, which highlights the characteristic group diethylstilbestrol phenol in the molecular structure of diethylstilbestrol. The diethylstilbestrol antibodies prepared had high recognition ability to diethylstilbestrol. The hapten and ovalbumin (OVA) carrier protein are coupled by active ester method (DCC, NHS).

[0077] b. Synthesis of Immunogen

[0078] Diethylstilbestrol and sodium chloroacetate are synthesized into a hapten through a halogen substitution reaction, and a spacer arm containing 2 carbon chains is connected to diethylstilbestrol, which highlights the characteristic group diethylstilbestrol phenol in the mole...

Embodiment 2

[0093] Embodiment 2 detects the formation of the ELISA kit of diethylstilbestrol

[0094] An enzyme-linked immunosorbent assay kit for detecting diethylstilbestrol was constructed to include the following components:

[0095](1) an enzyme-linked plate coated with diethylstilbestrol antigen;

[0096] (2) Goat anti-mouse anti-antibody labeled with horseradish peroxidase;

[0097] (3) Diethylstilbestrol mouse monoclonal antibody;

[0098] (4) 6 bottles of diethylstilbestrol 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;

[0099] (5) The substrate chromogenic solution A liquid is hydrogen peroxide, and the substrate chromogenic liquid B liquid is o-phenylenediamine;

[0100] (6) The stop solution is 2mol / L sulfuric acid buffer;

[0101] (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;

[0102] (8) The concentra...

Embodiment 3

[0103] Embodiment 3 detects the formation of the ELISA kit of diethylstilbestrol

[0104] An enzyme-linked immunosorbent assay kit for detecting diethylstilbestrol was constructed to include the following components:

[0105] (1) Enzyme-linked plate coated with goat anti-rabbit anti-antibody;

[0106] (2) diethylstilbestrol antigen labeled with alkaline phosphatase;

[0107] (3) Diethylstilbestrol rabbit polyclonal antibody;

[0108] (4) 6 bottles of diethylstilbestrol 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;

[0109] (5) The substrate chromogenic solution is p-nitrophosphate buffer;

[0110] (6) The stop solution is 2mol / L sodium hydroxide buffer solution;

[0111] (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;

[0112] (8) The concentrated complex solution is a phosphate buffer solution containing...

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Abstract

The invention provides an enzyme immune agent box for detecting atilen residual quantity of animal foodstuff which comprises: enzyme mark plate which coats atilen antigen or antibody, enzyme mark material, atilen peculiar antibody, atilen standard solution, 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 diethylstilbestrol in animal-derived food. Background technique [0002] Diethylstilbestrol (DES) is a synthetic non-steroidal estrogen, which is illegally used because it can promote protein synthesis and improve animal weight gain and feed efficiency. [0003] Studies have found that long-term intake of diethylstilbestrol drugs or foods containing diethylstilbestrol can lead to feminization, precocious puberty, and inhibition of bone and sperm development. In particular, these substances have obvious carcinogenic effects, which can lead to reproductive organ deformities in women and their female offspring and cancerous. In 2002, the Ministry of Agriculture issued the "List of Veterinary Drugs and Other Compounds Standardly Used in Food Animals", which clearly stipulates that diethylstilbestrol and its salts, esters, and preparations must not be used in food animals. [0004] At present,...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/52G01N33/535G01N33/543
Inventor 沈建忠何方洋冯才伟万宇平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 BEIJING WANGER BIOTECH
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