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Brucellosis indirect enzyme-linked immunosorbent assay antibody detection kit

An enzyme-linked immunosorbent adsorption and brucellosis technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of long reaction time, poor sensitivity and specificity, etc., and achieve the effect of simple operation and objective judgment of results

Inactive Publication Date: 2010-08-11
张晓艳
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, tiger red and test tube agglutination methods are used in our country. Their sensitivity and specificity are poor, and they are eliminated by international trade. The reaction time is long, and there is a certain contradiction with the actual detection and census work requirements in our country.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, the brucellosis indirect enzyme-linked immunosorbent assay antibody detection kit of the present invention, it is to utilize the purified lipopolysaccharide mixture of smooth type Brucella and rough type Brucella as antigen-coated microtiter plate, use S1119.3 Prepare positive control serum and standard weak positive serum from immunized healthy cattle, use healthy non-immune bovine serum as negative control serum, use enzyme-labeled receptor that binds to mammalian Fc segment-protein AG as enzyme-labeled conjugate, Equipped with serum diluent, washing solution, substrate solution A, substrate solution B, H 2 o 2 , stop solution, assembly composition.

[0044] The preparation method of the antigen-coated microtiter plate is as follows:

[0045] (1) Bacterial strains used to make antigens are B.abortus S1119.3 and B.abortus RB51 respectively; B.abortus S1119.3 strains can grow transparent on potato infusion agar after incubation at 37°C for 48 hours. , c...

Embodiment 2

[0086] Embodiment 2, the relevant problems of the brucellosis indirect enzyme-linked immunosorbent assay antibody detection kit of the present invention are explained as follows.

[0087] 1 strain selection

[0088] 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.

[0089] 2 Antigen pr...

Embodiment 3

[0111]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 F...

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Abstract

The invention aims to provide a brucellosis indirect enzyme-linked immunosorbent assay antibody detection kit which has moderate cost and short reaction time and is closely combined with the requirements of actual detection and general survey operations in China. The brucellosis indirect enzyme-linked immunosorbent assay antibody detection kit is assembled by using a purified lipopolysaccharide mixture of smooth type brucella and rough type brucella as antigen coated wells, using S1119.3 immune and healthy cattle to prepare positive control serum and standard weak positive serum, using healthy and non-immune cattle serum as negative control serum, using protein AG which is labeled by enzyme and is a receptor in combination with mammal Fc segment as enzyme labeling combo, and together taking serum diluent, a cleaning solution, a substrate solution A, a substrate solution B, H2O2 and a stopping solution as coordination. The invention establishes a rapid, sensitive, special and accurate method suitable for brucellosis diagnosis and epidemiological survey, has objective result judgment, simple and convenient operation, and is suitable for both large-scale screening tests and final confirmed diagnosis of the brucellosis.

Description

(1) Technical field [0001] The invention relates to medical biological product technology, in particular to a brucellosis indirect enzyme-linked immunosorbent assay 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 agglutina...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/535G01N33/543
Inventor 张晓艳
Owner 张晓艳
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