Method for detecting sulfanilamide medicine and special enzyme-linked immunoassay reagent kit thereof

A technology of enzyme-linked immunosorbent reagents and drugs, which is applied in the preparation methods of peptides, biochemical equipment and methods, and food testing, etc., and can solve problems such as cumbersome processes, unsuitable for on-site monitoring and screening of a large number of samples, and complex instruments and equipment

Inactive Publication Date: 2011-06-01
北京维德维康生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The chemical methods for the detection of sulfonamides include high performance liquid chromatography and liquid chromatography-tandem mass sp

Method used

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  • Method for detecting sulfanilamide medicine and special enzyme-linked immunoassay reagent kit thereof
  • Method for detecting sulfanilamide medicine and special enzyme-linked immunoassay reagent kit thereof
  • Method for detecting sulfanilamide medicine and special enzyme-linked immunoassay reagent kit thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Embodiment 1, sulfamethazine antigen, sulfamethazine antibody and preparation thereof

[0071] 1. Synthesis of sulfamethazine antigen

[0072] 1. Sulfamethazine immunogen

[0073] Sulfamethazine (SM 2 ) as the sulfamethazine hapten.

[0074] 1) Take 104mg sulfamethazine (SM 2 ), add 8mL 0.25mol L -1 sulfuric acid aqueous solution, placed in a 4 ° C refrigerator to form a solution I (containing SM 2 );

[0075] 2) 200 mg of human serum albumin (HSA) was dissolved in 8 mL of Na 2 CO 3 solution (pH = 10), placed in a refrigerator at 4°C to form solution II (containing HSA);

[0076] 3) 38mg NaNO 2 , add 2mL of pure water to obtain solution III (containing NaNO 2 );

[0077] 4) Add solution III (containing NaNO 2 ) slowly added to solution I (containing SM 2 ), during this process, solution I (containing SM 2 ) placed in crushed ice for about 15min to obtain solution IV (containing SM 2 -NaNO 2 mixture);

[0078] 5) Solution IV (containing SM 2 -NaNO 2 Mi...

Embodiment 2

[0100] Embodiment 2, composition and preparation of the ELISA kit for detecting sulfamethazine

[0101] 1. The enzyme-linked immunosorbent assay kit consists of the following materials:

[0102] 1. The sulfa drug coating former shown in formula II described in embodiment 1;

[0103] 2. ELISA plate;

[0104] 3. Sulfamethazine monoclonal antibody: secreted by hybridoma cell line SAs of sulfonamide monoclonal antibody hybridoma cell line with preservation number CGMCC No.3393;

[0105] 4. Sulfamethazine standard substance: Sulfamethazine, the concentration of the standard solution is 0μg / L, 0.5μg / L, 1.5μg / L, 4.5μg / L, 13.5μg / L and 40.5μg / L L;

[0106] Sulfamethazine standard was purchased from Sigma Aldrich; CAS number: 57-68-1; diluted with sample diluent to the above concentrations;

[0107] 5. Goat anti-mouse secondary antibody labeled with horseradish peroxidase;

[0108] 6. Substrate chromogenic solution: composed of liquid A and liquid B, liquid A is an aqueous solution...

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Abstract

The invention discloses a method for detecting a sulfanilamide medicine and a special enzyme-linked immunoassay reagent kit thereof. The enzyme-linked immunoassay reagent kit comprises a sulfanilamide medicine monoclonal antibody and the sulfanilamide medicine. The sulfanilamide medicine monoclonal antibody is secreted by the hybridoma cell line SAs of a sulfanilamide monoclonal antibody, whose preservation number is CGMCC No. 3393. In the enzyme-linked immunoassay reagent kit of the invention, an indirect competition ELISA (Enzyme-Linked Immuno Sorbent Assembly) method is mainly used for qualitatively or quantitatively detecting the content of the sulfanilamide medicine in products (especially milk, pork, chicken, eggs, honey, fish, shrimp and the like) eaten by animals or people. The reagent kit and the detection method of the invention have low requirements on the pretreatment of samples and simple pretreatment process of the samples and can detect mass samples quickly at the same time. By using the sulfanilamide medicine monoclonal antibody with high specificity, the detection method is convenient and simple, and the invention has the characteristics of high specificity, high sensibility, high precision, high accuracy and the like.

Description

technical field [0001] The invention relates to a method for detecting sulfa drugs and a special ELISA kit. Background technique [0002] Sulfonamides are artificially synthesized sulfonamide derivatives, mainly used for the prevention and treatment of bacterial infectious diseases. Representative drugs are: sulfamethazine, sulfadiazine, sulfamethoxazole, sulfamethoxine, sulfaquinoxaline, etc. Sulfonamide drugs can inhibit Gram-positive bacteria and some negative bacteria, and can treat a variety of bacterial infections. They are widely used in veterinary clinics to treat various livestock and poultry diseases infected by sensitive bacteria. [0003] Due to the long time of action and metabolism of sulfa drugs in the body, sulfa drugs ingested through any route may accumulate in the human body. When the accumulated concentration exceeds a certain value, it will cause damage to the human body. Short-term high-dose or long-term low-dose stimulation can cause acute or chroni...

Claims

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Application Information

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IPC IPC(8): C07K16/44C12N5/20C07K14/765C07K14/77C07K14/795C07K14/435G01N33/577G01N33/12G01N33/08G01N33/04G01N33/02C07K1/113
Inventor 江海洋吴小平王战辉徐飞史为民李娜王照鹏王进米铁军王亚辉
Owner 北京维德维康生物技术有限公司
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