Mink Aleutian disease virus antibody colloidal gold test strip and manufacturing method thereof

An Aleutian virus and detection test paper technology, which is applied in the field of mink Aleutian virus antibody colloidal gold detection test strip and its preparation, can solve the problems of complicated detection steps, time-consuming and the like, and achieves easy popularization, simple and quick operation, and high sensitivity. high effect

Active Publication Date: 2015-12-23
JL TEYAN BIOLOGICAL TECH LIMITED LIABILITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problem that the existing AD detection technology has cumbersome and time-consuming detection steps, requires professionals to operate and the detection mainly relies on laboratories, the present invention provides a colloidal gold detection test strip for mink Aleutian virus antibody and its preparation method

Method used

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  • Mink Aleutian disease virus antibody colloidal gold test strip and manufacturing method thereof
  • Mink Aleutian disease virus antibody colloidal gold test strip and manufacturing method thereof
  • Mink Aleutian disease virus antibody colloidal gold test strip and manufacturing method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0034] The preparation of embodiment 1 mink Aleutian virus (AMDV-G strain) cell antigen

[0035] Prepare mink Aleutian virus cell antigen by monolayer cell adsorption and inoculation method: CRFK subcultured cells are used for cell culture. After CRFK cells grow to a single layer, discard the original culture medium, add mink Aleutian virus, and incubate at 37°C for 1 hour , add cell culture medium containing 5% serum, culture statically at 37°C and observe the CRFK cell lesion every day, and harvest the cell culture medium when 85% to 90% of the CRFK cells have lesions, and freeze and thaw 3 times , put the cell culture virus liquid into a sterile centrifuge tube, lyse the cells and centrifuge at a rate of 10,000rpm / min for 2 hours to take the supernatant, and purify the virus by discontinuous sucrose density gradient centrifugation to obtain purified mink Aleutian virus cells antigen.

Embodiment 2

[0036] The preparation of embodiment 2 colloidal gold solution

[0037] Prepare colloidal gold solution by trisodium citrate reduction method: take 1ml of 1% chloroauric acid solution, add 99ml of ultrapure water to configure a final concentration of 0.01% chloroauric acid solution, heat to boiling, add 1% citric acid Trisodium 1ml and continue to heat, the solution turns from light yellow to blue-black and finally to wine red. After the color is stable, continue to heat for 5 minutes, cool at room temperature and store at 4°C for later use to obtain a colloidal gold solution. Take a small amount of colloidal gold particles and observe with a transmission electron microscope. The size of the colloidal gold particles is basically the same, the distribution is uniform, and the diameter is about 40nm.

Embodiment 3

[0038] Example 3 Preparation of Mink Aleutian Virus Cell Antigen and Colloidal Gold Coupling Marker (Gold Label Mink Aleutian Virus Cell Antigen)

[0039] (1) Stepwise dilution method determines the consumption ratio of mink Aleutian virus cell antigen and colloidal gold solution: adjust the pH value of colloidal gold solution with 0.1mol / L potassium carbonate solution to be 8.4, get 11 clean centrifuge tubes, numbered 1 to 11 tubes, add 1ml of colloidal gold solution to each tube; after the purified mink Aleutian virus cell antigen is diluted step by step (diluted from 5ug to 50ug, that is, the content of the purified mink Aleutian virus cell antigen is 5ug respectively , 10ug, 15ug, 20ug, 25ug, 30ug, 35ug, 40ug, 45ug, 50ug), according to the order of serial dilution, add to tube No. 2 to tube No. 11, and mix with the colloidal gold solution in the centrifuge tube for 5 minutes Then add 1ml of 10% sodium chloride solution to tube No. 2 to tube No. 11, mix well, let stand at r...

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Abstract

The invention provides a mink Aleutian disease virus antibody colloidal gold test strip and a manufacturing method thereof and relates to the technical field of colloidal gold test paper strips. The problem that an existing AD detection technology comprises complicated detection steps, consumes time, needs a specialized person to operate and mainly depends on a laboratory during detection is solved. The test strip comprises a PVC base plate, a sample pad, a gold labeled conjugate pad, a nitrocellulose membrane provided with a detection line and a quality control line and a water absorption pad, wherein the sample pad, the gold labeled conjugate pad, the nitrocellulose membrane and the water absorption pad are sequentially pasted on the PVC base plate. The gold labeled conjugate pad is coated with a purified mink Aleutian disease virus cell antigen and colloidal gold coupling marker and a mouse IgG and colloidal gold coupling marker, the detection line on the nitrocellulose membrane is coated with a purified mink Aleutian disease virus cell antigen, and the quality control line on the nitrocellulose membrane is coated with goat anti-mouse IgG. The test strip has the advantages that the test strip is simple and quick to operate, good in specificity and high in sensibility, a detection result is clear and easy to judge, no instruments or equipment is needed.

Description

technical field [0001] The invention relates to the technical field of colloidal gold detection test strips, in particular to a colloidal gold detection test strip for mink Aleutian virus antibody and a preparation method thereof. Background technique [0002] Mink Aleutian disease (Aleutian Disease, AD) is caused by mink Aleutian virus (Aleutian Mink Disease Virus, AMDV), a chronic infectious disease of immune system disorder and abnormal metabolism. Mink Aleutian disease can seriously affect the fur quality of mink, damage its fertility, reduce the survival rate, and bring huge economic losses to the mink industry. It is known as one of the three major diseases of fur animals. Prevention and control of the mink Aleutian disease. [0003] At present, the conventional AD detection technology mainly adopts convective immunoelectrophoresis (CIEP), but the detection steps of this method are cumbersome, time-consuming, require professionals to operate, and mainly rely on labora...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56983
Inventor 王振军王春霞倪佳张晶王萃瑜吴威陈立志
Owner JL TEYAN BIOLOGICAL TECH LIMITED LIABILITY
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