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ELISA kit for detecting chloramphenicols in animal derived food

A technology for detecting animals and chloramphenicol, applied in the field of immunological detection, can solve the problems of high cost, cumbersome processing and measurement operations, and restrictions on popularization and use, and achieves simple sample pretreatment process, no radioisotope pollution, and reagent storage time. long effect

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

Due to the cumbersome sample pretreatment and high cost of instrument analysis methods such as high performance liquid chromatography and gas chromatography, the popularization and use of them are limited.

Method used

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  • ELISA kit for detecting chloramphenicols in animal derived food
  • ELISA kit for detecting chloramphenicols in animal derived food

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] The preparation of embodiment 1 kit components

[0087] 1. Synthesis of hapten

[0088] The alcoholic hydroxyl group in the molecular structure of chloramphenicol is acylated with glutaric anhydride to obtain the hapten of chloramphenicol.

[0089] 2. Antigen synthesis

[0090] a. Dissolve 2 g of chloramphenicol hapten in 20 ml of 0.5 M sodium hydroxide solution.

[0091] b. Dissolve 1.5 g of carbodiimide in 5 ml of pure water and add it to the hapten solution and stir at room temperature for 2 hours.

[0092] c. Dissolve 20 g of carrier protein rabbit serum albumin (RSA) or thyroid protein (BCG) in 75 ml of pH9.6 carbonate buffer.

[0093] d. Add the carrier protein dropwise to the hapten and stir overnight at 4°C.

[0094] e. Dialyze the reacted artificial antigen against 0.1M phosphate buffer for 7 days, and change the solution 3 to 4 times a day. Finally, the antigen is concentrated or lyophilized for storage.

[0095] 3. Preparation of enzyme-labeled anti-ant...

Embodiment 2

[0110] Embodiment 2 detects the formation of the ELISA kit of chloramphenicol drugs

[0111] Set up the ELISA kit for detecting chloramphenicol so that it includes the following components:

[0112] (1) ELISA plates coated with chloramphenicol antigens;

[0113] (2) Chloramphenicol mouse monoclonal antibody working solution with a protein concentration of 0.5 μg / L;

[0114] (3) Goat anti-mouse anti-antibody labeled with horseradish peroxidase;

[0115] (4) 6 bottles of chloramphenicol standard solution, the concentrations were 0 μg / L, 0.05 μg / L, 0.15 μg / L, 0.45 μg / L, 1.35 μg / L, 4.05 μg / L;

[0116] (5) The substrate chromogenic solution A liquid is hydrogen peroxide, and the substrate chromogenic liquid B liquid is o-phenylenediamine;

[0117] (6) The concentrated washing solution is a phosphate buffer containing 0.8% Tween 20 and 1‰ sodium azide preservative;

[0118] (7) The concentrated complex solution is a phosphate buffer containing 5‰N,N'-dimethylformamide.

[0119]...

Embodiment 3

[0120] Embodiment 3 detects the formation of the ELISA kit of chloramphenicol drugs

[0121] Set up the ELISA kit for detecting chloramphenicol so that it includes the following components:

[0122] (1) A microtiter plate coated with goat anti-mouse anti-antibody;

[0123] (2) Chloramphenicol mouse monoclonal antibody working solution with a protein concentration of 5.0 μg / L;

[0124] (3) chloramphenicol antigen labeled with alkaline phosphatase;

[0125] (4) 6 bottles of chloramphenicol standard solution, the concentrations were 0 μg / L, 0.05 μg / L, 0.15 μg / L, 0.45 μg / L, 1.35 μg / L, 4.05 μg / L;

[0126] (5) The substrate solution is p-nitrophosphate buffer;

[0127] (6) The concentrated washing solution is a phosphate buffer containing 1.2% Tween 20 and 1‰ sodium azide preservative;

[0128] (7) The concentrated complex solution is a phosphate buffer containing 5‰N,N'-dimethylformamide.

[0129] (8) The stop solution is 2mol / L sodium hydroxide solution.

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Abstract

The invention provides an enzyme immune agent box for detecting chloromycetin drugs of animal foodstuff which comprises: enzyme mark plate which coats chloromycetin drugs antigen or antibody, chloromycetin drugs mouse monoclonal antibody compression solution, chloromycetin drugs standard solution, enzyme mark material, 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, in particular to an ELISA kit for detecting chloramphenicol drugs in animal-derived food and a detection method thereof. Background technique [0002] Chloramphenicol (Chloramphenicol, CAP) is widely used antibiotics, once played an important role in the control and treatment of livestock and poultry diseases. However, because chloramphenicol residues in food have serious side effects on the human body and can cause human diseases such as aplastic anemia and agranulocytosis, developed countries such as Europe and the United States have banned or strictly prohibited their use. According to the No. 235 document of the Ministry of Agriculture of my country, the residue limit of chloramphenicol in food of animal origin shall not be detected. When detected by enzyme-linked immunoassay method and high-performance liquid chromatography (HPLC), the detection limit is required to be 1ng / kg. Due to...

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 BEIJING WANGER BIOTECH
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