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Method for preparing germs polyclonal antibody of sugarcane ratoon stunting disease

A polyclonal antibody, dwarf disease technology, applied in the preparation method of peptide, chemical instrument and method, organic chemistry and other directions, to achieve the effect of high purity and titer, low cost and high specificity

Inactive Publication Date: 2009-10-21
GUANGZHOU SUGARCANE IND RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has the characteristics of a large number of detectable samples, simple operation, and high accuracy, but it is limited by the need for specific antibodies
At present, there is no report on the preparation of this antibody. Therefore, it is urgent to prepare the antibody of this pathogen to meet the needs of RSD pathogen diagnosis in sugarcane scientific research and production.

Method used

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  • Method for preparing germs polyclonal antibody of sugarcane ratoon stunting disease

Examples

Experimental program
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Effect test

Embodiment 1

[0030] A preparation method of polyclonal antibody against sugarcane ratoon stunting disease pathogen is carried out according to the following steps:

[0031] (1) Isolation and cultivation of pathogenic bacteria to obtain antigens;

[0032] (1.1) Obtaining sugarcane juice: From November to December, in fine weather, cut off the mature cane stems of susceptible varieties, and after removing surface impurities, clean the surface with detergent, dry, and wipe with 70% alcohol Disinfect the surface; cut the cane stalk into 5-10cm length, then cut it longitudinally, and use pliers to directly take the cane juice. The cane juice is placed in a 50ml centrifuge tube.

[0033] (1.2) Sugarcane juice filtration and low-speed centrifugation: filter the sugarcane juice with filter paper, transfer the filtered cane juice to a new centrifuge tube, centrifuge at low speed (2000rpm) for 5 minutes, and transfer the supernatant to a new centrifuge tube , Repeat the above centrifugation step once.

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Embodiment 2

[0075] A preparation method of polyclonal antibody against sugarcane ratoon stunting disease pathogen. The steps are the same as those in Example 1, and the main differences are:

[0076] The SC medium used in step (1) (1.3) pathogen cultivation step and (1.4) pathogen subculture purification step is a modified SC medium: cornmeal agar(15-18)g / L+Bacto- agar(3-5)g / L+Bacto-peptone(6-9)g / L+Bovine hemin chloride15mg / L+MgSO 4 ·7H 2 O(0.1-0.2)g / L+K 2 HPO 4 13ml(0.1M storage solution) / L+KH 2 PO 4 87ml(0.1M storage solution) / L; Glucose(0.5-2)g / L+Cysteine(1-2)g / L+Bovine serum albumin 2g / L, PH6.5-8.0;

[0077] The liquid SC medium used in the step (1) (1.5) expanding the cultivation of pathogenic bacteria to prepare the antigen is a modified liquid SC medium: Bacto-peptone(6-9)g / L+Bovine heminchloride 15mg / L+MgSO 4 ·7H 2 O(0.1-0.2)g / L+K 2 HPO 4 13ml(0.1M storage solution) / L+KH 2 PO 4 87ml(0.1M storage solution) / L; Glucose(0.5-2)g / L+Cysteine(1-2)g / L+Bovine serum a lbumin 2g / L, PH6.5-8.0.

[...

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Abstract

The invention relates to the technical field for preventing and curing sugarcane ratoon stunting disease, providing a method for preparing germs polyclonal antibody of sugarcane ratoon stunting disease, comprising the following steps: (1) culturing pathogenic germs by means of isolation to obtain antigen; (2) mixing the antigen with equivalent Freund complete adjuvant, emulsifying, and immunizing rabbit; (3) mixing the antigen with equivalent Freund incomplete adjuvant, emulsifying, and again immunizing rabbit, repeating the immunizing process several times, after measuring of titer meets requirement, collecting blood from the heart, isolating antiserum; and (4) purifying the antiserum to obtain the polyclonal antibody. In the invention, the germs of the sugarcane perennial root dwarfing is isolated from sugarcane juice, thus the polyclonal antibody which is obtained thereby can specifically identify the germs of the sugarcane ratoon stunting disease in the sugarcane juice. The germs polyclonal antibody of the sugarcane ratoon stunting disease has the advantages of high specificity, high purity and high titer, can be preserved for a long time, and has important practical significance in measuring the pathogenic germs to diagnose the sugarcane ratoon stunting disease (RSD) by means of immunology in the scientific research and the production of the sugarcane.

Description

Technical field [0001] The invention relates to the technical field of polyclonal antibody preparation, in particular to a preparation method of polyclonal antibody against sugarcane ratoon stunting disease bacteria. Background technique [0002] Sugarcane ratoon stunting disease (RSD) is one of the most important bacterial diseases in sugarcane production. The pathogen Lxx is very small and parasitizes in the xylem ducts of sugarcane. Because the disease has no obvious external and internal symptoms, and it is easy to be confused with the symptoms caused by drought, and it is mainly spread through germ-carrying seed stems and cutting tools, which leads to unintentional transmission, causing the disease to spread, and harming sugarcane production. great. The disease usually causes sugarcane yield to be reduced by 12-37%. Under drought conditions, the yield is reduced by up to 60%, and the ratoon is weakened. Even disease-free healthy seedlings are 100% susceptible after being use...

Claims

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Application Information

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IPC IPC(8): C07K16/12C07K1/36C07K1/34C07K1/30
Inventor 陈健文刘睿齐永文何方洋万宇平余厚美邓海华潘永保沈万宽杨湛端
Owner GUANGZHOU SUGARCANE IND RES INST
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