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EHV-1 serum antibody detection kit as well as preparation method and application

A technology of EHV-1 and detection kit, which is applied in the field of virus serum antibody detection, can solve the problems that are not suitable for ordinary outdoor technicians, can not be used to detect EHV-1, and the detection time is long, so as to achieve wide practicability and application value , good reactogenicity, simple operation effect

Inactive Publication Date: 2016-07-20
XINJIANG AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

All of the above methods need to be tested in the laboratory, which has the characteristics of long detection time and cumbersome process. It is more suitable for laboratory diagnosis, but not suitable for ordinary outdoor technicians, and cannot be applied to the detection of EHV-1 in large quantities of samples in grassroots units.
[0004] Therefore, by developing a fast, simple, and easy-to-carry diagnostic device, a method that can quickly detect EHV-1 can be provided, which can solve the problem of long detection time and cumbersome process, and can make a timely diagnosis of EHV-1 infection. Accurate diagnosis has extensive practical significance for the field of biological detection

Method used

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  • EHV-1 serum antibody detection kit as well as preparation method and application
  • EHV-1 serum antibody detection kit as well as preparation method and application
  • EHV-1 serum antibody detection kit as well as preparation method and application

Examples

Experimental program
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Effect test

Embodiment 1

[0043] Example 1: EHV-1 Serum Antibody Detection Kit

[0044] The present invention specifically provides a kind of EHV-1 serum antibody detection kit, EHV-1 serum antibody detection kit comprises, box cover, NC film, water-absorbing material and box bottom, and kit adopts by plastic small box as shell, prefabricated NC The membrane and water-absorbing material constitute the inner core of the kit. The box body is a cuboid structure with a length of 3.0cm, a width of 3.0cm, and a height of 0.5cm. There is a sample hole with a diameter of 0.5cm in the center of the box cover, and a layer of water-absorbing material is provided at the bottom of the box. The NC membrane is located vertically above the water-absorbing material, wherein the NC membrane is respectively coated with purified EHV-1gD inclusion body protein and porcine immunoglobulin IgG.

[0045] When using the EHV-1 serum antibody detection kit provided by the present invention, the NC membrane is first coated with th...

Embodiment 2

[0047] Example 2: Preparation of EHV-1 Serum Antibody Detection Kit

[0048] The present invention specifically provides a kind of EHV-1 serum antibody detection kit, and specific production steps are as follows:

[0049] (1) Preparation of NC membrane: The NC membrane is coated with purified EHV-1gD inclusion body protein and porcine immunoglobulin IgG respectively, and the gD inclusion body protein and porcine immunoglobulin IgG are both added dropwise to the NC membrane The method of natural drying is used for coating, the purified EHV-1gD inclusion body protein is used as the detection antigen, and porcine immunoglobulin IgG is used as the quality inspection protein, and the purified EHV-1gD inclusion body protein is transferred into the pET28a-gD prokaryotic expression plasmid The BL21 (DE3) Escherichia coli was induced and expressed by 0.9mmol / L IPTG at 25°C for 6h, and the cells were ultrasonically lysed and the lysate was washed with PBS. The washing method was to coll...

Embodiment 3

[0051] Example 3: Application of EHV-1 Serum Antibody Detection Kit

[0052] Adopt the application of the kit that above-mentioned embodiment one and embodiment two provide in EHV-1 serum antibody detection, concrete application method is as follows:

[0053] (1) Before using the kit, place the kit device horizontally, add 2 drops of PBS blocking solution containing 2% BSA and 0.05% Tween20 to the NC membrane of the kit, and the dropping amount is 100ul for blocking;

[0054] (2) After the blocking solution on the NC membrane has completely penetrated into the NC membrane, add 1 drop of the tested horse serum, and the dropping amount is 50ul;

[0055] (3) After the horse serum has completely penetrated into the NC membrane, add gold standard protein A dilution solution, and the dropping amount is 100ul;

[0056] (4) After the diluent has completely infiltrated into the NC membrane, add 2 drops of PBST for washing, the dropping amount is 100ul, repeat 3 times;

[0057] (5) Ob...

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Abstract

The invention discloses an EHV-1 serum antibody detection kit as well as a preparation method and application. The EHV-1 serum antibody detection kit takes a plastic small box as a shell, and a kit inner core is composed of a prefabricated NC film and filter paper; the kit shell is of a cuboid structure; the NC film is coated with purified EHV-1 gD inclusion body protein and pig immunoglobulin IgG respectively; the purified EHV-1 gD inclusion body protein is used as a detection antigen; the pig immunoglobulin IgG is used as quality inspection protein; the purified EHV-1 gD inclusion body protein and the pig immunoglobulin IgG are assembled according to a sequence of a shell cover, the NC film, the filter paper and a shell bottom; a gold label SPA is used as a second antibody, PBS is used as a coating solution, and PBS is used as washing liquid; and an EHV-1 antibody condition in a horse blood serum sample is detected by adopting an antigen and antibody specific combining principle. The method is rapid, visible and simple to operate, is suitable for basic staff to use and has a wide application value.

Description

technical field [0001] The present invention relates to the technical field of biological detection, specifically, the present invention relates to a virus serum antibody detection method and the technical field of application thereof. Background technique [0002] Equinerhinopneumonia (Equinerhinopneumonitis, ER) is a general term for several highly contagious diseases of equine animals. (EHV-4), EHV-1, also known as equine abortion virus, mainly causes abortion in mares. In areas where a large number of horses are raised, the positive rate of horse EHV-1 infection is about 40%, and more than half of the horses in some areas are serologically positive. Sick horses and recovered infected horses are the main source of infection for this disease. The clinical symptoms are mild in adult horses and severe in foals, with clinical manifestations of fever, catarrhal inflammation of the upper respiratory mucosa, and leukopenia. The course of the disease is about 1 week, and the fo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/569G01N33/558
CPCG01N33/558G01N33/56994G01N33/6854G01N2469/20
Inventor 王世民张艳楠
Owner XINJIANG AGRI UNIV
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