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Hapten, preparation method and application for detection of Daowenling content

A rice blast spirit and hapten technology, which is applied in chemical instruments and methods, measuring devices, biological tests, etc., can solve the problems of lack of hapten and other problems, and achieve the effects of strong specificity, simple detection operation and convenient use.

Active Publication Date: 2022-07-01
YUNNAN TOBACCO QUALITY SUPERVISION MONITORING STATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The invention provides a hapten for detecting the content of rice blastling, a preparation method and its application, which are used to solve the lack of antibodies with specific reactions to the ingredients of rice blastling and the technology of hapten for synthesizing antibodies existing in the prior art question

Method used

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  • Hapten, preparation method and application for detection of Daowenling content
  • Hapten, preparation method and application for detection of Daowenling content
  • Hapten, preparation method and application for detection of Daowenling content

Examples

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Effect test

Embodiment 1

[0062] The preparation of the above-mentioned hapten includes the following steps:

[0063] Take 1.88g of diisopropyl malonate, add 20mL of carbon disulfide, stir at room temperature, mix well, add 13mL of 2mol / L KOH solution, stir at room temperature for 1h, then add 1.56g of 3,4-dichlorobutyric acid, and add 2mol / L KOH solution 5mL, tetrabutylammonium bromide 3.8g, acetonitrile 80mL, heated and refluxed for 4h, the reaction was stopped, rotary evaporated, acetonitrile and carbon disulfide were removed, 80mL of water was added, 6mol / L HCl was added to adjust the pH to 6, and acetic acid was added. Ethyl ester was extracted three times with 100 mL × 3, the organic phases were combined, washed with water, dried over anhydrous sodium sulfate and evaporated to dryness to obtain a yellow oil, which was applied to a silica gel column and eluted with a 3:1 petroleum ether-ethyl acetate mixed solution by volume. After purification, 0.76 g of acetic acid-Diawenling was obtained, whic...

Embodiment 2

[0065] The preparation of an antigen for the detection of rice blast spirit content includes the following steps:

[0066] Take 12.9 mg of the hapten prepared in Example 1, add 1 mL of tetrahydrofuran to dissolve and clarify, add 8.5 mg of N-hydroxysuccinimide (NHS), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide 14.8 mg of imine hydrochloride (EDC), fully dissolved and mixed, and reacted at room temperature for 2 h to obtain hapten activation solution A; take 50 mg of bovine serum albumin (BSA), add 0.1 mol / L CB9.1 8 mL to dissolve, and obtain Solution B; add solution A to solution B dropwise, react at room temperature for 4 hours, stop the reaction, dialyze and purify it with 0.02mol / L PBS buffer for 3 days, change the solution 3 times a day, and centrifuge to obtain a compound with bovine serum albumin. Combined rice blastling artificial antigen, aliquot, and store at -20°C.

Embodiment 3

[0068] The preparation of an antigen for the detection of rice blast spirit content includes the following steps:

[0069] Take 7.7 mg of rice blast spirit hapten prepared in Example 1, add 1 mL of dimethyl sulfoxide (DMSO) to dissolve and clarify, add 5.1 mg of NHS and 9.7 mg of EDC, fully dissolve and mix, and react at room temperature for 2 hours to obtain a hapten activation solution A solution; take ovalbumin (OVA) 50mg, add 0.1mol / L CB9.1 7mL to dissolve, obtain B solution; add A solution dropwise to B solution, react at room temperature for 4h, stop the reaction, use 0.02mol / L L PBS buffer was dialyzed and purified for 3 days, the medium was changed three times a day, and centrifuged to obtain the artificial antigen of rice blast spirit conjugated with ovalbumin, which was packaged and stored at -20°C.

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Abstract

The invention discloses a hapten for detecting the content of rice blast spirit, a preparation method and application thereof, and in particular relates to a hapten for enzyme-linked immunoassay detection of rice blast spirit. The invention also discloses a preparation method of the hapten and its application. Application in antibodies and kits. The rapid detection product of the enzyme-linked immunosorbent assay kit based on the rice blast hapten is easy to use, low in detection cost, efficient, accurate, fast, and can detect a large number of samples at the same time. It is suitable for agricultural products, especially the rice blast in tobacco. On-site monitoring and screening of large samples.

Description

technical field [0001] The invention relates to the technical field of pesticide residue detection, in particular to a hapten for detecting the content of rice blast spirit, a preparation method and an application thereof. Background technique [0002] Isoprothiolane, also known as isopropyl sulfur ring, Fuji No. 1, chemical name 1,3-dithiolane-2-ylidene malonate diisopropyl ester, is a highly efficient systemic fungicide. Its bactericidal spectrum is narrow, and it is a special medicine for the control of rice blast, especially for panicle neck blast. It is also effective against rice seedling blast and sclerotinia sclerotiorum, and can also treat rice planthoppers and leafhoppers when used in a large area, and has a regulating effect on plant growth. However, with the large-scale application of rice blast spirit, it may pollute the environment, increase its residual amount in agricultural products, and pose a potential threat to human health. It is now necessary to carry...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D339/06C07K14/765C07K14/77C07K14/795C07K16/44G01N33/53
CPCC07D339/06C07K14/765C07K14/77C07K14/795C07K16/44G01N33/5308
Inventor 王春琼陈丹张燕秦世春李苓蔡洁云张轲张晓伟彭丽娟孙浩巍杨艺敏张庆刚
Owner YUNNAN TOBACCO QUALITY SUPERVISION MONITORING STATION
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