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Reagent kit for detecting o-allyl phenol and its special antibody

A technology of o-allylphenol and kits, which is applied in the field of kits for detecting o-allylphenol and its special antibodies, can solve the problems of difficult measurement of a large number of samples, expensive experimental equipment, high cost of reagents, etc., and achieve a high derivatization rate High, high detection limit, good effect

Inactive Publication Date: 2009-08-05
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have the following main limitations: long sample pretreatment time, expensive experimental equipment, high cost of reagents, etc.; there are certain difficulties in the determination of a large number of samples, especially in the rapid detection of pesticide residues in fruits, these methods are powerless

Method used

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  • Reagent kit for detecting o-allyl phenol and its special antibody
  • Reagent kit for detecting o-allyl phenol and its special antibody
  • Reagent kit for detecting o-allyl phenol and its special antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1, the synthesis of o-allylphenol antigen (CAH antigen)

[0049] The schematic diagram of the synthesis of o-allylphenol antigen is as follows figure 1 shown.

[0050] 1. Synthesis of o-allylphenol immune antigen (CAH-bovine serum albumin (BSA))

[0051]1. Diazotization:

[0052] (1) Take 30 mg of o-allylphenol, dissolve in 1 ml of methanol, dilute to 5 mL with distilled water, and stir magnetically at room temperature.

[0053] (2) Take 40mg of p-aminobenzoic acid, dissolve it in 1ml, 1M NaNO 2 1M HCl was added dropwise, and the reaction was terminated when the starch potassium iodide test paper turned blue.

[0054] (3) Slowly add the solution obtained in step (2) dropwise to the solution obtained in step (1) under constant stirring, and the reaction is complete after 12 hours at 4°C. When the pH value is adjusted to 2-3, a large amount of brick red precipitates out , centrifuge the precipitate; dissolve the precipitate in distilled water, adjust the p...

Embodiment 2

[0072] Example 2. Preparation of ortho-allylphenol monoclonal antibody and its identification

[0073] 1. Preparation of monoclonal antibody

[0074] (1) Establishment of hybridoma cell lines

[0075] 1. Animal immunity

[0076] The immunogen CAH-BSA was dissolved in 0.5 mL of normal saline, and an equal volume of complete adjuvant was added to make an immunogen emulsion, and 6-week-old female Balb / c mice were immunized. The second immunization was performed 4 weeks later, and the third immunization was performed two weeks later, using an immunogen emulsifier prepared by adding an equal volume of incomplete adjuvant. On the 10th day after the third immunization, the antibody titer in the mouse serum was measured by indirect ELISA, and the one with the higher titer was subjected to the next step of cell fusion.

[0077] 2. Cell Fusion

[0078] Spleen of Balb / c mice immunized with CAH-BSA was aseptically obtained to prepare spleen cell suspension. Draw separately containing...

Embodiment 3

[0120] Example 3, Preparation of o-allylphenol polyclonal antibody

[0121] (1) Balb / C female mice aged 8-10 weeks were used as experimental animals. Experimental immunization dose: 0.25-2.0 mg / kg for basic immunization, and 0.5-2.0 mg / kg for booster immunization.

[0122] (2) Basic immunization: Dilute the CAH-BSA immune antigen obtained in Example 1 with sterile water, add an equal volume of Freund's complete adjuvant after filtration through a sterile filter, and fully emulsify until dripped into water without spreading. The emulsified immune antigen was injected subcutaneously into the animal at multiple points on the back, and 0.2 mL was injected.

[0123] (3) Booster immunization: use incomplete Freund's adjuvant to emulsify the immune antigen, the method is the same as (2). Booster immunization is carried out every 3-4 weeks, and the abdominal cavity and the back are injected alternately. From the third immunization, on the 8th-10th day after each immunization, blood ...

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Abstract

The invention discloses a reagent kit for detecting o-allyl phenol and a special antibody thereof. The reagent kit comprises a monoclonal antibody of the o-allyl phenol or a polyclonal antibody of the o-allyl phenol, wherein the monoclonal antibody of the o-allyl phenol is generated by the excretion of mouse hybridoma cell line mAb225# with the preserving number of CGMCC No.2905. The enzyme linked immune reagent kit for detecting o-allyl phenol and a method for detecting the o-allyl phenol can be used for detecting the residual quantity of the o-allyl phenol in farm products such as plant tissues, has the advantages of simple sample preliminary treatment course, simple and convenient operation, low cost, good specificity, high sensitivity and precision, and the like, can monitor on site and is suitable for screening a plurality of samples, therefore, the method for detecting the o-allyl phenol and the special reagent kit thereof can exert an important function in detecting the residual o-allyl phenol chemical in the farm products.

Description

technical field [0001] The invention relates to a kit for detecting o-allylphenol and its special antibody. Background technique [0002] 2-propenylphenol (2-propenylphenol, trade name Yinguo) is a new type of agricultural fungicide developed by the Agricultural Biomimetic Application Engineering Research Center of Shandong Province. Botrytis), which is now mass-produced. It is based on the chemical structure of the bactericidal and antibacterial active compounds in Ginkgo biloba. After the structure is simplified, it is obtained through artificial chemical synthesis. The molecular weight is 134, and the molecular formula is C 9 h 10 O. Field efficacy tests have proved that o-propenylphenol can effectively control strawberry powdery mildew, tomato botrytis, apple rot, corn leaf spot and other diseases. However, if the amount of fungicide is too much, it will remain in agricultural products and affect people's health. [0003] At present, the analysis methods of o-allylp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N30/02C07K16/44C07K14/765C07K14/77C12N5/20
Inventor 李健强夏源源王保民曹永松刘西莉罗来鑫刘鹏飞
Owner CHINA AGRI UNIV
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