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ELISA (enzyme-linked immuno sorbent assay) detection method of rabbit swainsonine-resistant antibody, and kit

A technology of swainsonine and detection method, which is applied in the field of immunology, can solve the problems of ELISA detection kit research reports and patent applications without rabbit anti-swainsonine antibody, and achieve high precision and optimized detection Conditions, the effect of easy operation

Inactive Publication Date: 2013-04-17
TARIM UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

And so far, there is no research report and patent application on the ELISA detection kit of rabbit anti-swainsonine antibody

Method used

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  • ELISA (enzyme-linked immuno sorbent assay) detection method of rabbit swainsonine-resistant antibody, and kit
  • ELISA (enzyme-linked immuno sorbent assay) detection method of rabbit swainsonine-resistant antibody, and kit
  • ELISA (enzyme-linked immuno sorbent assay) detection method of rabbit swainsonine-resistant antibody, and kit

Examples

Experimental program
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preparation example Construction

[0031] (1) Preparation of coated antigen: first react bromoacetic acid with N-hydroxysuccinimide to form an active ester, and then synthesize the active ester with SW to form N-carboxymethyl swainsonine, and then react with N-carboxymethyl Swainsonin is condensed with OVA, and freeze-dried after dialysis is SW-OVA. The chemical reaction formula is as follows: Figure 1-Figure 3 shown.

[0032] (2) Preparation of positive serum: use swainsonine artificial antigen SW-BSA to immunize rabbits, use Freund's complete adjuvant (FAC) to immunize rabbits for basic immunization, and perform first immunization after 10 days, use FAC and normal saline The mixed solution (volume ratio 1:1) fully emulsified the prepared SW-BSA; the first and second booster immunizations were emulsified with a mixture of Freund's incomplete adjuvant (FAI) and normal saline (volume ratio 1:1) ; For the third booster immunization, physiological saline was used to dissolve SW-BSA, and 10 days after the third b...

Embodiment 1

[0035] Example 1 uses swainsonine artificial antigen to immunize rabbits, which is made by a method consisting of the following steps:

[0036] 1. Preparation of swainsonine artificial antigen: first react bromoacetic acid with N-hydroxysuccinimide to form an active ester, then synthesize the active ester with SW to form N-carboxymethyl swainsonine, and then react with N-carboxysuccinimide Methyl swainsonine is condensed with BSA, freeze-dried after dialysis to obtain SW-BSA, and its chemical reaction formula is as followsfigure 1 , figure 2 , Figure 4 shown.

[0037] 2. Rabbits immunized with swainsonine artificial antigen: Rabbits were randomly divided into negative control group (1) and SW-BSA immunized group (6), and Freund's complete adjuvant (FAC) was used for immunization group. For immunization, use the mixture of FAC and normal saline (volume ratio 1:1) to fully emulsify the prepared SW-BSA; for the first and second booster immunization, use Freund's incomplete ad...

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Abstract

The invention discloses an ELISA (enzyme-linked immuno sorbent assay) detection method of a rabbit swainsonine-resistant antibody, and a kit. The indirect ELISA method is adopted for detecting the rabbit serum swainsonine-resistant antibody, and comprises the following steps of: coating; sealing; adding to-be-detected serum and positive and negative serum; adding IgG-HRP (immunoglobulin G-Horse Radish Peroxidase); adding substrate solution; and suspending. Through the implementation of the method, the immune effect of a rabbit is judged rapidly, sensitively and accurately; and the ELISA detection method is simple and feasible, and is suitable for scientific research institutions and livestock raising systems of grass roots.

Description

technical field [0001] The invention belongs to the technical field of immunology, and relates to an ELISA detection method and a kit for rabbit anti-swainsonine antibody. Background technique [0002] Locoweed is a collective name for poisonous plants of the leguminosae genus Oxytropis and Astragalas, and the poisoning disease caused by it is called Locosim or Locodisease. . Locoweeds in my country are mainly distributed in Inner Mongolia, Ningxia, Gansu, Shaanxi, Qinghai, Xinjiang, Tibet and other provinces. The distribution area has exceeded 11 million hm 2 , accounting for about 2.8% of the total grassland area in the country, accounting for 3.3% of the grassland area in the west, and spreading at a rate of 3.5% every year. Locoweed has become the main poisonous weed that endangers the sustainable development of grassland animal husbandry in southwest and northwest pastoral areas of my country. Swainsonine (SW) is one of the main toxic components of locoweed, which i...

Claims

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Application Information

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IPC IPC(8): G01N33/53
Inventor 王帅马春晖胡建军席琳乔王连群陈根元
Owner TARIM UNIV
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