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Immunological magnetic bead for melon bacterial fruit spot disease and its preparation method

A technology of immune magnetic beads and bacteria, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of no report on the identification of plant pathogenic bacteria, and achieve the effect of improving specificity and sensitivity, high sensitivity, and strong specificity

Inactive Publication Date: 2007-12-12
XINJIANG XIYU IND GROUP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] Adsorption media are widely used in the fields of biochemistry and molecular biology to separate and purify biomolecules such as nucleic acids and proteins, but there is no report on the identification of plant pathogens

Method used

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  • Immunological magnetic bead for melon bacterial fruit spot disease and its preparation method
  • Immunological magnetic bead for melon bacterial fruit spot disease and its preparation method
  • Immunological magnetic bead for melon bacterial fruit spot disease and its preparation method

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Experimental program
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Effect test

Embodiment 1

[0026] 1. Preparation of BFB immune antibody;

[0027] Expand the culture of the standard BFB pathogen and collect the bacteria, extract the protein of the bacteria by ultrasonic peak, make the adjuvant BFB antigen, immunize rabbits, wait until the titer reaches 1:64, bloodletting to obtain anti-BFB serum, salt out with ammonium sulfate, FpLc protease A column purification to obtain BFB antibody IgG.

[0028] 2. Making BFB immunomagnetic beads

[0029] After the above-mentioned BFB antibody IgG is coupled with a solid-phase carrier, it becomes BFB immunomagnetic beads. The production route is as follows:

[0030] (1), the choice of solid phase carrier. The carrier is a polysaccharide polymer, SephadexCellulose or Sephadex-agrose can be selected.

[0031] (2) The solid-phase carrier is cross-linked with the ion-exchange group diethylaminoethyl to form an ion-exchange solid-phase carrier. The response is as follows:

[0032]

[0033] (3) Activation of the ion-exchange soli...

Embodiment 2

[0037] Embodiment 2: the preparation method of the immune magnetic beads of melon bacterial fruit rot spot of the present invention is:

[0038] 1. Cultivate the standard BFB pathogen and collect the bacteria, extract the protein of the bacteria by ultrasonic crushing, make an adjuvant BFB antigen, immunize rabbits, and wait until the titer reaches 1:64, bloodletting to obtain anti-BFB serum, and use ammonium sulfate Analysis, FpLc protease A column purification to obtain BFB antibody IgG.

[0039] 2. Take 5 grams of solid carrier Sephadex or Cellulose with anion-exchange gene, add 250ml of sterile water, and let it swell overnight.

[0040] 3. Put 1ml of the above gel into a small chromatography column, elute 10ml with pH6.8, 0.01mol / L TB containing 0.4mol / L NaCl, and then use pH6.8, 0.01mol / L TB solution to balance wash Remove 20ml.

[0041] 4. Take 0.5ml of the purified BFB antibody containing 5mg of protein and add it to the above-mentioned small chromatography column. A...

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Abstract

The invention relates to the making of agent for BFB bacteria of melons. It comprises the making of BFB immune antibody and BFB immune magnetic pearl, selecting non-solutble inertia as the solid carrier to exchange base chain with the ions to form ion exchange solid carrier with large quantity of exchangeable negative ion, activating the ion exchange solid carrier with the BFB immune antibody to get the BFB immune magnetic pearl. It uses the immune theory to couple the antibody with the inertia organic molecules, to establish the BFB quick inspection, quick, convenient, high in sensitivity, fine in special feature, improved in feature and sensitivity.

Description

1. Technical field [0001] The invention belongs to a specific reagent for detecting melon bacterial fruit rot caused by oat acidophilus subsp. Immunomagnetic beads and preparation methods. 2. Background technology [0002] Acidovorax avenae subsp.citrulli, or Aac for short, can cause Bacterial fruit blotch of cucurbits, or BFB for short, which is a worldwide quarantine pest. In particular, the above-mentioned diseases are the three types of dangerous diseases stipulated in the "List of Dangerous Diseases, Insects and Weeds for Imported Plant Quarantine of the People's Republic of my country" published by our country. [0003] The incidence area of ​​melon bacterial fruit rot is expanding. The symptoms at the seedling stage of the disease often manifest as large black lesions without any water-soaked shape, which makes field diagnosis very difficult. The isolation and culture diagnosis of pathogenic bacteria takes 10-14 days, and the procedure is cumbersome. When the carrie...

Claims

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Application Information

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IPC IPC(8): G01N33/547G01N33/546G01N33/531G01N33/569
Inventor 喻梅辉张红马光玉王叶筠周志成丁建军穆全昌姜明张兴平
Owner XINJIANG XIYU IND GROUP
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