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Brucellosis indirect enzyme-linked immunosorbent assay milk liquid antibody reagent kit

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

Active Publication Date: 2013-04-10
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

[0027] Embodiment 1, brucellosis indirect enzyme-linked immunosorbent assay milk antibody detection kit of the present invention, it is to utilize smooth type Brucella (B.abortus S1119.3) and rough type Brucella (B.abortus The purified lipopolysaccharide mixture of RB51) was used as an antigen-coated microtiter plate, the positive control milk was prepared by immunizing healthy cattle with S19 vaccine, the negative control milk was prepared by healthy non-immunized cattle, and the receptor bound to the Fc segment of mammals was labeled with enzyme Body - protein AG as enzyme labeling conjugate, with diluent, washing solution, substrate solution A, substrate solution B, H 2 o 2 , stop solution, assembly composition. The present invention also has the following technical characteristics:

[0028] 1. the preparation method of described antigen-coated microtiter plate is as follows:

[0029] (1) Coating: Under sterile conditions, suspend the two purified lyophilized antigens wi...

Embodiment 2

[0052] Embodiment 2, with regard to the relevant problem of brucellosis indirect enzyme-linked immunosorbent assay milk antibody detection kit of the present invention is described as follows:

[0053] 1 strain selection

[0054] Brucella bovis S1119.3 is a traditional strain for extracting antigen. It is derived from USDA of the United States. Because S1119.3 is not easy to transform into rough type, it is a strain approved by the International Organization for Animal Health (OIE) to extract lipopolysaccharide from smooth type bacteria. RB51 is a variant strain of Brucella bovis rough type. As a vaccine strain of Brucella bovis, it is also an international general strain for extracting rough type bacterial lipopolysaccharide. The mixture of two bacterial lipopolysaccharides is used as an antigen to detect brucellosis antibodies, the purpose is to detect all serotypes of brucella antibodies, improve the detection rate of the detection kit, and avoid missed detection.

[0055...

Embodiment 3

[0077] 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 indirect enzyme-linked immunosorbent assay milk liquid antibody reagent kit which meets the demand of epidemic prevention of animals in our country and is quick, sensitive, specific and steady. The reagent kit is assembled by utilizing a purified lipopolysaccharide mixture of smooth bacterium burgeri and rough bacterium burgeri as an antigen coated ELIAS plate, using an immune healthy cow to prepare a positive control milk liquid, using a healthy non-immune cow to prepare a negative control milk liquid, taking a receptor-protein AG which is labeled by enzyme and is combined with Fc segments of mammals as an enzyme labeled conjugate, and is matched with a diluent, a washing liquid, a substrate solution A, a substrate solution B, H2O2, and a stop buffer. The brucellosis indirect enzyme-linked immunosorbent assay milk liquid antibody reagent kit establishes a quick, sensitive, specific and accurate inspection method which is suitable for brucellosis diagnosis and epidemiological investigation, and meets the demand of the epidemic prevention of the animals in our country.

Description

(1) Technical field [0001] The invention relates to an immunological testing method, in particular to a brucellosis indirect ELISA milk antibody 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 be...

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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