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Rabies antibody gold immunochromatography assay testing indicator paper and preparation technique

A technology for immunochromatographic detection and rabies virus, which is applied to measurement devices, analytical materials, instruments, etc., can solve the problem of no simple and fast detection, etc., and achieve the effects of easy operation, simple preparation method, and easy interpretation.

Inactive Publication Date: 2007-08-15
MILITARY VETERINARY RES INST PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, for the prevention of canine rabies, the method of injecting rabies vaccine is adopted, but whether effective antibodies are produced in the dog body after vaccination, there is no simple and rapid detection method and reagent

Method used

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  • Rabies antibody gold immunochromatography assay testing indicator paper and preparation technique

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Experimental program
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Effect test

Embodiment 1

[0026] 1. Preparation of goat-anti-dog IgG: Goat-anti-dog IgG made by immunizing sheep with purified canine immunoglobulin as an antigen, and the antibody is only directed against canine immunoglobulin. Use this antibody to mark colloidal gold to make gold-labeled antibody, which can combine with canine immunoglobulin in the sample to be tested, so use this antibody to mark colloidal gold and spray it on the binding pad.

[0027] 2. Preparation of rabbit anti-goat IgG: This antibody is made by immunizing rabbits with sheep immunoglobulin as an antigen, and extracting the antibody globulin in the immune rabbit serum. Used on the quality control line of the NC membrane, it can be combined with the gold-labeled antibody to detect the validity of the test strip.

[0028] 3. Preparation of rabies virus antigen: use attenuated ERA strain of rabies virus vaccine to infect monolayer Vero cells, collect the supernatant after lesion as the first-generation virus, and then infect monolay...

Embodiment 2

[0030] The production process of test strips:

[0031] 1. Preparation of sheep anti-canine IgG gold-labeled antibody (1) Preparation of colloidal gold: take concentration of 0.01% chloroauric acid (HAuCl 4) solution 100mL heating and boiling, quickly add a certain amount of 1% trisodium citrate aqueous solution, and then keep boiling for 10-15min, the colloidal gold solution with a gold particle size of 20 nanometers can be obtained; (2) colloidal gold-labeled sheep anti-canine IgG : Use 0.1M potassium carbonate (K 2 CO 3 ) solution to adjust the pH to 8.2, add goat anti-canine IgG at 2-18ug / ml, after mixing and stirring, add bovine serum albumin (BSA) at 1g / 100ml to the solution, and let stand at 4°C for 2-4 hours. The above colloidal gold solution was centrifuged at 2000r / min for 20 minutes, and the precipitate was removed to obtain the supernatant. Then the supernatant was centrifuged at 10,000 r / min for 40 minutes to obtain a precipitate. Resuspend the precipitate with...

Embodiment 3

[0037] (1) The bonding pad is made of glass fiber membrane, which is produced by Millipore Company of the United States. The bonding pad is treated with 10mMpH8.2 PBS+3%BSA+0.1%Triton X-100+1%PVP-40000 pretreatment solution and soaked for 30 minutes Afterwards, vacuum-dry at 37°C, spray colloidal gold-labeled goat anti-dog IgG and then dry. The upper part of the binding pad is connected with the sample pad, and the lower part is connected with the NC membrane;

[0038] (2) NC membrane is a kind of material specially used in the preparation of colloidal gold test strips. It is produced by Millipore Company of the United States. Two lines are sprayed on the NC membrane, and one is a detection line (purified rabies virus antigen or antigen expressed by genetic engineering) ), the other line is a quality control line (rabbit anti-goat IgG), the NC membrane is pasted in the middle of the backing, and the two ends are respectively connected with the binding pad and the water-absorbin...

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Abstract

This invention provides one dog rabies virus antigen glue gold immune chromatography test paper and its process, which comprises the following steps: according to antigen antibody immune combination basic principle to label the sheep IgG by glue gold label covering on the glass fiber film as combination pad; covering the purification rabies virus antigen and gene engineer expressed antigen and rabit IgG covering on the NC film as test line and quality control line; when testing the antibody, forming antibody combination to expose red band.

Description

Technical field: [0001] The invention relates to a test strip for rapid detection of specific rabies virus effective protective antibody in dogs, in particular provides a test strip for colloidal gold immunochromatography detection of canine rabies virus antibody and a preparation process, and belongs to the technical field of animal epidemic detection. Background technique: [0002] Rabies was recorded as early as the fourth century BC. It is a direct contact and viral infectious disease caused by rabies virus. All warm-blooded animals are susceptible to it. It is an important zoonotic disease that endangers human health. According to statistics, 35,000-50,000 patients died of rabies every year in the world. From 1950 to 2005, the total number of rabies deaths in my country was about 102,280. The total number of morbidity and death is second only to India, ranking second in the world. Rabies dogs, cats, and certain wild animals are the most important sources of infection fo...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/558G01N33/543G01N33/532
Inventor 夏咸柱黄耕杨松涛王承宇高玉伟侯小强高明华王铁成薛琳
Owner MILITARY VETERINARY RES INST PLA MILITARY MEDICAL ACAD OF SCI
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