New-generation brucellosis antibody competition enzyme-linked immunosorbent adsorption test detection kit

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

Active Publication Date: 2009-11-18
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

[0034]Embodiment 1, a new generation of brucellosis antibody competition ELISA detection kit of the present invention, it is to utilize the lipopolysaccharide of smooth type Brucella as antigen-coated microtiter plate, immunization with inactivated bacterial liquid Prepare positive control serum and standard weak positive serum from healthy cows or artificially infected healthy cows, use healthy non-immune cow serum as negative control serum, use new monoclonal antibodies as competing antibodies, and anti-Fc receptor protein AG enzyme markers as secondary antibodies , prepare serum diluent, washing solution, substrate solution A, substrate solution B, H 2 o 2 , stop solution, assembly composition.

[0035] The new generation of brucellosis antibody competition ELISA detection kit of the present invention also has the following technical characteristics:

[0036] 1. The antigen-coated microtiter plate is prepared in this way:

[0037] (1) Coating: under sterile conditions, s...

Embodiment 2

[0072] Embodiment 2, with regard to the related problems of the new generation brucellosis antibody competition enzyme-linked immunosorbent assay detection kit of the present invention, do as follows:

[0073] 1 About strain selection and standards

[0074] 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.

[0075] 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.

[0076] 2 About antigen preparation

[0077] 2.1 Control of Bacterial Propagation Conditions S1119.3 is an international sta...

Embodiment 3

[0092] 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 new-generation brucellosis antibody competition enzyme-linked immunosorbent adsorption test detection kit which meets animal epidemic prevention requirements in China, is sensitive and specific and can accurately diagnose brucellosis. In the invention, the lipopolysaccharide of smooth type brucella is used as an antigen-coated enzyme-labeled plate, inactivated bacilli liquid is used for immunizing or artificially infecting a healthy cattle to prepare positive control sera and standard weak positive sera, healthy non-immune cattle sera are used as negative control sera, a novel monoclonal antibody is used as a competition antibody, the protein AG enzyme-labeled substance of an anti-Fc receptor is used as an assistant antibody, serum diluent, a washing solution, a substrate solution A, a substrate solution B, H2O2 and a stop solution are prepared, and all raw materials are assembled to form the kit. The invention is a competition ELISA kit which is sensitive and specific and can accurately diagnose brucellosis. The invention meets animal epidemic prevention requirements in China, and carries out last diagnosis on the brucellosis by means of a laboratory diagnosis technique.

Description

(1) Technical field [0001] The invention relates to immunological testing technology, in particular to a new generation of brucellosis antibody competition enzyme-linked immunosorbent assay 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, s...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569G01N33/531
Inventor 张晓艳
Owner 哈尔滨平河生物技术有限公司
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