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ELISA kit for detecting furazolidone metabolites and detection method thereof

A technology of furazolidone and metabolites, which is applied in the field of enzyme-linked immunosorbent assay kits, and can solve the problems of cumbersome operation, high instrument cost, and complicated processing

Active Publication Date: 2008-06-25
BEIJING WANGER BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, liquid chromatography (LC) and high-performance liquid chromatography (HPLC) are commonly used methods for detecting furazolidone residues in aquatic products and animal husbandry. These methods are complicated in sample pretreatment, high in instrument cost, and cumbersome in operation, and are only suitable for confirmation of samples. analysis, not suitable for screening of large numbers of samples

Method used

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  • ELISA kit for detecting furazolidone metabolites and detection method thereof
  • ELISA kit for detecting furazolidone metabolites and detection method thereof
  • ELISA kit for detecting furazolidone metabolites and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1 Preparation of ELISA Kit Components for Detecting Furazolidone Metabolites

[0076] 1. Antigen Synthesis

[0077] a. Synthesis of Coating Progen

[0078] The furazolidone metabolite is synthesized by a derivative method to synthesize a furazolidone metabolite derivative hapten, and then the hapten is obtained by coupling the hapten with a thyroid protein carrier protein by a mixed anhydride method through a diazotization reaction.

[0079] b. Synthesis of Immunogen

[0080] The furazolidone metabolite is synthesized by a derivative method to synthesize a furazolidone metabolite derivative hapten, and then the hapten is obtained by coupling the hapten through a diazotization reaction and an ovalbumin carrier protein by a mixed anhydride method.

[0081] 2. Preparation of mouse monoclonal antibody against furazolidone metabolite derivatives

[0082] a. Animal immunity

[0083] Balb / c mice were used as immunized animals, and furazolidone metabolite derivative...

Embodiment 2

[0094] Example 2 Construction of an enzyme-linked immunosorbent assay kit for detecting furazolidone metabolites

[0095] An enzyme-linked immunosorbent assay kit for detecting furazolidone metabolites was constructed to include the following components:

[0096] (1) Enzyme-linked plates coated with furazolidone metabolite derivative antigens;

[0097] (2) Goat anti-mouse anti-antibody labeled with horseradish peroxidase;

[0098] (3) Furazolidone metabolite derivative mouse monoclonal antibody;

[0099] (4) 6 bottles of furazolidone metabolite 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;

[0100] (5) The substrate chromogenic solution A liquid is hydrogen peroxide, and the substrate chromogenic liquid B liquid is o-phenylenediamine;

[0101] (6) The stop solution is 2mol / L sulfuric acid buffer;

[0102] (7) The concentrated washing solution is pH 7.4, containing 0.8% Tween 20 and 1‰ sodium azide (NaN 3 ) phosphate...

Embodiment 3

[0105] Example 3 Construction of an enzyme-linked immunosorbent assay kit for detecting furazolidone metabolites

[0106] An enzyme-linked immunosorbent assay kit for detecting furazolidone metabolites was constructed to include the following components:

[0107] (1) Enzyme-linked plate coated with goat anti-rabbit anti-antibody;

[0108] (2) Furazolidone metabolite derivative antigen labeled with alkaline phosphatase;

[0109] (3) Furazolidone metabolite derivative rabbit polyclonal antibody;

[0110] (4) 6 bottles of furazolidone metabolite derivative 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;

[0111] (5) Substrate chromogenic solution p-nitrophosphate buffer;

[0112] (6) The stop solution is 2mol / L sodium hydroxide buffer solution;

[0113] (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;

[0114] ...

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Abstract

The invention provides an enzyme immune agent box for detecting furazolidone metabolism product of animal foodstuff which comprises: enzyme mark plate which coats furazolidone metabolism product derivative antibody or the enzyme mark plate which coats the antibody, enzyme mark material, furazolidone metabolism product derivative peculiar antibody, furazolidone metabolism product derivative 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 furazolidone metabolites in animal-derived foods. Background technique [0002] Furazolidone is a synthetic broad-spectrum antibacterial drug with stable efficacy, which can inhibit or kill a variety of Gram-positive and negative bacteria, and has certain effects on some protozoa and fungi. Due to its high efficiency and low cost, furazolidone has been widely used in animal husbandry and aquaculture. [0003] However, the residues of furazolidone in animal-derived foods can be transmitted to humans through the food chain, and long-term intake can cause various diseases. According to the report of the Food and Agriculture Organization of the United Nations (FAO) and the World Health Organization (WHO) Joint Expert Committee on Food Additives, furazolidone and its metabolites are carcinogenic and prone to drug-resistant strains, which can easily cause teratogenicity. my country listed furaz...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/543G01N33/52G01N33/535
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
Owner BEIJING WANGER BIOTECH
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