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Brucellosis antibody competitive enzyme-linked immunosorbent assay reagent kit

An enzyme-linked immunosorbent assay and brucellosis technology, applied in the field of immunological testing, can solve the problems of backward technology and high false positives

Active Publication Date: 2013-04-24
哈尔滨平河生物技术有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the tiger red plate and test tube agglutination methods currently used in my country are eliminated by international trade, the technology is backward, and the false positives are relatively high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1, the brucellosis antibody competition enzyme-linked immunosorbent assay detection kit of the present invention, it is to utilize the lipopolysaccharide of smooth type brucella to make the antigen-coated microtiter plate, immunize healthy cattle with inactivated bacterial liquid Or artificially infect healthy cattle to prepare positive control serum and standard weak positive serum, use healthy non-immune bovine serum as negative control serum, use monoclonal antibody as competing antibody, enzyme-labeled secondary antibody (goat anti-mouse IgG), prepare serum diluent, Washing solution, substrate solution A, substrate solution B, H 2 o 2 , stop solution, assembly composition.

[0037] The brucellosis antibody competition ELISA detection kit of the present invention also has the following technical characteristics:

[0038] 1. The antigen-coated microtiter plate is prepared in this way:

[0039] (1) Coating: under sterile conditions, suspend the freeze-dri...

Embodiment 2

[0066] Embodiment 2, with regard to the related problems of the brucellosis antibody competition enzyme-linked immunosorbent assay detection kit of the present invention, be explained as follows:

[0067] 1 About strain selection and standards

[0068] Strain selection and seed algebra Many Brucella have been isolated at home and abroad, with different serotypes. At present, the Brucella that I keep and use is the bovine type S1119.3 strain, which is an international standard strain for preparing brucellosis detection antigens. This strain is easy to cultivate and is not prone to gross changes. Freeze-dried and stored at -70°C, the shelf life is at least 10 years.

[0069] According to the genetic stability study of the bacterium, the structure of the SLPS antigenic site is more stable after 10 generations of continuous passage of the bacterium.

[0070] 2 About antigen preparation

[0071] 2.1 Control of Bacterial Propagation Conditions S1119.3 is an international standard...

Embodiment 3

[0086] Example 3, the primary binding test for diagnosing brucellosis, including fluorescence polarization detection and enzyme-linked immunosorbent assay. The fluorescence polarization method is a homogeneous method that does not need to remove unbound reagents. The operation speed is fast, and the result can be obtained within two minutes, which can eliminate some vaccine reactions. It can be used not only in laboratory testing, but also in pastures and fields, and is suitable for the detection of brucellosis in wild animals. FPA has high sensitivity and is an excellent screening test. Relatively inexpensive and as accurate as other primary binding assays. This type of assay is also amenable to automation. It works by measuring the change in molecular reactivity caused by the reaction of antibodies with soluble antigen in the test sample. This response is measured if the antibody binds the antigen and the rate of rotation of the antigen decreases. The basic principle of ...

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Abstract

The invention aims to provide a brucellosis antibody competitive enzyme-linked immunosorbent assay reagent kit which meets the demand of epidemic prevention of animals in our country, is sensitive and specific, and can accurately diagnose brucellosis. The reagent kit is assembled by utilizing lipopolysaccharide of smooth brucella as an antigen coated ELIAS plate, using an inactivated bacterial liquid to immunize a healthy cow or artificially infect the healthy cow to prepare positive control serum and standard weak-positive serum, using serum of a healthy non-immune cow as negative control serum, using a monoclonal antibody as a competitive antibody, namely an ELIAS secondary antibody (goat anti-rat IgG), and preparing a serum diluent, a washing liquid, a substrate solution A, a substrate solution B, H2O2, and a stop buffer. The brucellosis antibody competitive enzyme-linked immunosorbent assay reagent kit is a competitive ELISA reagent kit which is sensitive and specific and can accurately diagnose the brucellosis. The reagent kit meets the demand of the epidemic prevention of the animals in our country, and performs final definite diagnosis on the brucellosis by means of laboratory diagnosis technology.

Description

(1) Technical field [0001] The invention relates to an immunological testing technology, in particular to a brucellosis antibody competition ELISA detection kit. (2) Background technology [0002] Immunological detection methods are a series of experimental methods designed by applying immunological theory to detect antigens, antibodies, immune cells and their secreted cytokines. With the interpenetration of disciplines, the scope of immunology has been continuously expanded, and new immunological detection methods have emerged in an endless stream. The scope of application of immunological methods is also expanding, not only becoming an important method for the diagnosis of various clinical diseases, but also providing convenience for the research of many disciplines. [0003] When the antigen meets the corresponding antibody, specific binding can occur, and under the influence of external conditions, certain reaction phenomena, such as agglutination or precipitation, can ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/535G01N33/543
Inventor 张晓艳
Owner 哈尔滨平河生物技术有限公司
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