Flow-through Aleutian mink disease virus antigen antibody detection kit, preparation method thereof and application thereof

A technology of Aleutian virus and detection kit, which is applied to the detection kit for antigen and antibody of Aleutian virus in mink and its preparation field, can solve the problem that detection steps are cumbersome, time-consuming, hinder the popularization and promotion of Aleutian disease detection in mink, etc. problem, to achieve the effect of easy promotion, simple and fast use, and low manufacturing cost

Active Publication Date: 2019-10-25
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[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 laboratories. These disadvantages have seriously hindered the popularization of mink Aleutian disease detection. and promotion

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  • Flow-through Aleutian mink disease virus antigen antibody detection kit, preparation method thereof and application thereof
  • Flow-through Aleutian mink disease virus antigen antibody detection kit, preparation method thereof and application thereof

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preparation example Construction

[0050] The invention provides a preparation method of a through-flow mink Aleutian virus antigen antibody detection kit, comprising the following steps:

[0051] a) The mink Aleutian virus monoclonal antibody A1 is mixed and reacted with the colloidal gold solution, and the solid reactant is collected, and then sequentially sealed and concentrated to obtain the mink Aleutian virus monoclonal antibody diagnostic solution;

[0052] b) The mink Aleutian virus monoclonal antibody B1, the standard mink Aleutian virus and the quality control antibody were spotted and coated on the nitrocellulose membrane to form T1 spots, T2 spots and quality control spots respectively.

[0053] The steps of the method are simple and easy to operate, can be produced industrially, have good repeatability, and the produced product has stable performance. It should be noted that the coating positions of mink Aleutian virus monoclonal antibody B1, mink Aleutian virus and quality control antibody are dif...

Embodiment 1

[0067] Embodiment 1 Preparation of mink Aleutian virus monoclonal antibody A1, B1

[0068] (1) Using the hybridoma cell fusion method, female Balb / c mice aged 6-8 weeks were selected, and according to a specific immunization scheme, the mice were immunized 5 times multi-point subcutaneously and on the foot pads, and 3 times after the last immunization Splenocytes from immunized mice were fused with cultured myeloma cells, and cell lines capable of secreting specific mink Aleutian virus monoclonal antibodies were screened out and expanded to prepare mink Aleutian virus monoclonal antibodies A1 and B1;

[0069] (2) Specific immunization scheme: For the first immunization, the mice were mixed and emulsified with complete Freund's adjuvant and purified ADV at 1:1, and the mice were immunized at multiple points. Each mouse was immunized at a total of 5-8 points, with an interval of 3 weeks after the first immunization; for the second immunization, the mice were mixed and emulsified...

Embodiment 2

[0071] Embodiment 2 Preparation of mink Aleutian virus monoclonal antibody diagnostic fluid

[0072] (1) Mink Aleutian virus monoclonal antibody A1 is mixed with a colloidal gold solution with a particle size of 20-30nm, the mixing ratio is 30 μg-40 μg:1mL, and mink Aleutian virus monoclonal antibody A1 is added dropwise according to this dosage ratio, and then Add bovine serum albumin at a final concentration of 5%, and after stirring, obtain a conjugated marker of mink Aleutian virus monoclonal antibody A1 and colloidal gold, which is stored at a final concentration of 5% BSA, 10% sucrose, 0.2 MTris and 0.02% sodium azide, the buffer system with a pH value of 8.0-8.5 is ready for use;

[0073] (2) The colloidal gold solution adopts the preparation method of a high-power gold solution: get 1ml of 0.02% chloroauric acid solution, after heating to boiling, add 1% trisodium citrate 3ml and continue heating, the solution turns from light yellow to blue-black at last It is orange...

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Abstract

The invention relates to the field of animal disease detection and particularly relates to a flow-through Aleutian mink disease virus antigen and antibody detection kit, a preparation method thereof and application thereof. The kit provided by the invention comprises an Aleutian mink disease virus monoclonal antibody diagnostic solution and test paper, and the test paper comprises a nitrocellulosemembrane with detection points and a quality control point. The Aleutian mink virus monoclonal antibody diagnostic solution contains an Aleutian mink disease virus monoclonal antibody A1 marked by acolloidal gold, the detection points comprises a T1 point and a T2 point, the T1 point is coated with an Aleutian mink disease virus monoclonal antibody B1, the T2 point is coated with a standard Aleutian mink virus, and quality control point is coated with a quality control antibody. The kit has the function of simultaneously detecting the Aleutian mink virus antigen and antibody and can detect whether a sample is infected with the Aleutian mink virus and whether the Aleutian virus antibody is produced.

Description

technical field [0001] The invention relates to the field of animal epidemic detection, in particular to a through-flow mink Aleutian virus antigen antibody detection kit and a preparation method and application thereof. Background technique [0002] Mink Aleutian disease (Aleutian Disease, AD) is caused by mink Aleutian virus (Aleutian MinkDisease 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 laboratories. The...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/577
CPCG01N33/56983G01N33/577
Inventor 王振军罗国良程悦宁郭利易立冯二凯程世鹏
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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