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
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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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