ELISA kit for detecting avermectins and detection method thereof

An enzyme-linked immunosorbent reagent, abamectin technology, applied in the direction of biological testing, measuring devices, material inspection products, etc. Complicated and other problems, to achieve the effect of simple and easy test method, high specificity and simple processing process

Inactive Publication Date: 2006-05-03
贵州勤邦食品安全科学技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The detection by the above method has the defects of complex equipment, sample pretreatment and cumbersome measurement operation, it is...

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  • ELISA kit for detecting avermectins and detection method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] The preparation of embodiment 1 kit components

[0063] 1. Antigen Synthesis

[0064] a. Synthesis of hapten

[0065] Abamectin was synthesized by dibromosuccinic acid method and glutaric anhydride method to synthesize abamectin drug hapten.

[0066] b. Immunogen synthesis

[0067] The abamectin drug hapten and human serum albumin are coupled by a mixed anhydride method to obtain an immunogen.

[0068] The preparation process of the immunogen: take 2 g of abamectin drug hapten and dissolve it in 30 ml, 50% N, N-dimethylformamide solution, then take 0.5 ml of isobutyl chloroformate and dissolve it in 5 ml of anhydrous di Add it to the hapten solution and stir it at room temperature for 4 hours, take 32g of carrier protein and dissolve it in 70ml pH9.6 carbonate buffer, then add human serum albumin dropwise to the hapten and stir overnight at 4°C. The reacted artificial antigen was dialyzed against 0.2M phosphate buffer for 7 days, and the solution was changed 3 to 4 ...

Embodiment 2

[0081] Embodiment 2 detects the formation of the ELISA kit of avermectins

[0082] Set up the enzyme-linked immunosorbent assay kit that detects abamectin class drug, make it comprise following components:

[0083] (1) A microtiter plate coated with goat anti-mouse anti-antibody;

[0084] (2) Abamectin antigens labeled with horseradish peroxidase;

[0085] (3) Abamectin antibodies with a protein concentration of 0.5 μg / L;

[0086] (4) 6 bottles of ivermectin 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;

[0087] (5) The substrate chromogenic solution is composed of A liquid and B liquid, the substrate chromogenic liquid A liquid is carbamide peroxide, and the substrate chromogenic liquid B liquid is tetramethylbenzidine;

[0088] (6) The stop solution is 2mol / L hydrochloric acid;

[0089] (7) The concentrated washing solution contains 0.01M ~ 0.05M phosphate and 1‰ sodium azide (NaN 3 ) buffer;

[0090] (8) The con...

Embodiment 3

[0091] Embodiment 3 detects the formation of the ELISA kit of avermectins

[0092] Set up the enzyme-linked immunosorbent assay kit that detects abamectin class drug, make it comprise following components:

[0093] (1) A microtiter plate coated with goat anti-mouse anti-antibody;

[0094] (2) extracting the avermectin antigen labeled with alkaline phosphatase with bacteria;

[0095] (3) Abamectin antibody with a protein concentration of 5.0 μg / L;

[0096] (4) 6 bottles of ivermectin 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;

[0097] (5) The substrate chromogenic solution is p-nitrophosphate buffer;

[0098] (6) The stop solution is 2mol / L sodium hydroxide;

[0099] (7) The concentrated washing solution contains 0.01M ~ 0.05M phosphate and 1‰ sodium azide (NaN 3 ) buffer;

[0100] (8) The concentrated diluent is 0.1M phosphate buffer containing 1% casein.

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Abstract

The invention provides an enzyme immune agent box for detecting avermectin drugs which comprises: enzyme mark plate which coats sheep-resistance mouse-resistance antibody, enzyme mark antigen, avermectin drugs antibody, avermectin standard solution, base material color developing solution, compression cleaning liquid, ending solution and compression diluting 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 abamectin drugs in animal tissues. Background technique [0002] Avermectins (AVMS) belong to macrolide antibiotics, commercialized avermectins are a mixture of the two most active homosystems among eight compounds with very similar chemical structures, Avermectins The dihydrogen reduction product of ivermectin is another important antiparasitic drug. [0003] Abamectin has a strong inhibitory effect on the vast majority of parasitic nematodes and arthropods with important economic value. It is a major breakthrough in the research of antiparasitic drugs in the past ten years. At present, abamectins have begun large-scale production and application. [0004] However, the excessive use of abamectin drugs makes them highly residual in animal-derived foods, causing the human body to develop drug resistance and causing certain harm to the human body. Therefore, it has been used in limited qua...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/52G01N33/535G01N33/543
Inventor 沈建忠何方洋史为民丁双阳张素霞万宇平冯才伟吴小平冯才茂汪善良李军赵正苗张照亮罗晓琴孙倩
Owner 贵州勤邦食品安全科学技术有限公司
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