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Animal brucella antigen colloidal gold test paper film detection reagent kit

A technology of colloidal gold test paper and Brucella, which is applied in the field of animal epidemic diagnosis, can solve the problems of time-consuming detection, prone to false positives, laborious and other problems, and achieve the effect of rapid detection

Inactive Publication Date: 2007-10-17
MILITARY VETERINARY RES INST PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

They all have the following deficiencies. First, it is difficult to avoid cross-reaction between animal Brucella and other bacteria (such as: Yersinia O9, Escherichia coli O157); second, traditional detection methods are time-consuming and laborious. And prone to false positives; third, traditional diagnostic methods cannot distinguish the types of brucellosis

Method used

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  • Animal brucella antigen colloidal gold test paper film detection reagent kit
  • Animal brucella antigen colloidal gold test paper film detection reagent kit

Examples

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Effect test

Embodiment 1

[0031] 1. Extraction and identification of animal Brucella LPS and O chain

[0032] Two strains of Brucella glabrata, B.abortus 544A and B.melitensis 16M were selected, and the LPS and O chains of the above two strains were extracted by hot phenol method and cold phenol method respectively. After purification by SephadexG-50 chromatography column, The content is 1.21-2.31mg / mL, and the purity is 96.3%-97.2%. The purified LPS and O chain were identified, and the antigenicity of LPS and O chain of two strains of Brucella glabrata was tested.

[0033] 2. Preparation and identification of Brucella species-specific monoclonal antibodies

[0034] B. melitensis 16M and B. abortus 544A cells were used to immunize Balb / C mice, and purified LPS and O-chain antigens were used as detection antigens to produce species-specific monoclonal antibodies 4B8 and 2D10 against A and M antigens, and purified After potency is 1×10 6 Above, the monoclonal antibody subtypes are IgG1, IgG3 and IgG2b...

Embodiment 2

[0038] 1. Preparation of colloidal gold and selection of optimum particle size

[0039] Prepare 20nm, 25nm, 30nm, 40nm colloidal gold particles by sodium citrate reduction method and tannic acid-sodium citrate reduction method, observe the dispersion and uniformity under the electron microscope, select the most suitable colloidal gold particle size of 30nm, 40nm for labeling .

[0040] 2. Determination of the optimal labeling pH of the animal Brucella gold standard probe

[0041] Using colloidal gold gradient method and O value curve method, determine the optimal pH of colloidal gold-labeled monoclonal antibody IgG. The optimal standard pH for this test is 8.2-9.0.

[0042] 3. Determination of the optimal labeling amount of the animal Brucella gold standard probe

[0043]Using the colloidal gold gradient method and the O value curve method, determine the optimal amount of colloidal gold-labeled monoclonal antibody IgG. The optimal amount of labeling in this test is 20-60 μ...

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Abstract

The invention discloses an animal brucella antigen colloidal gold test paper film detection kit, which bases on immunology basic principle of antigen and antibody specific combination, utilizes colloidal gold immunity chromatography technique and silver dye reinforcement technique, uses combined golden yellow staphylococcus A protein (SPA) as gold marking antibody, uses high purity brucella LPS as peridium antigen, and utilizes goat anti-bovine IgG as quality control line to prepare colloidal gold test paper tape. The invention can accurately differentiate species of animal brucella and can synchronously detect animal brucella disease of cattle, sheep and pig species with no crossing reaction and low false negative proportion, negative and positive coincidence ratio of the invention and conventional separation and culture is 100%, detection result can be obtained in 5 min, and the kit need no assistant complicated detection instrument.

Description

technical field [0001] The invention discloses an animal Brucella antigen colloidal gold test paper film detection kit, which is used for the diagnosis of brucellosis and belongs to the technical field of animal epidemic diagnosis. Background technique [0002] Brucella is a Gram-negative, specific intracellular parasite that infects many species of animals and humans. At present, Brucella is divided into 6 species according to the pathogenicity and host, namely, Brucella sheep, Brucella bovis, Brucella pig, Brucella sylvania, and Brucella ovis. bacteria, Brucella canis. Among them, brucella of cattle, sheep, and pigs are widely prevalent in the world, which is extremely harmful to animal husbandry. More importantly, they can cause human infection and threaten public health. Therefore, the research on the diagnosis of brucellosis has always been concerned by countries all over the world. Traditional serological diagnostic methods for diagnosing animal brucellosis include:...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/569G01N33/558
Inventor 王兴龙李晓艳唐景峰梅建军
Owner MILITARY VETERINARY RES INST PLA MILITARY MEDICAL ACAD OF SCI
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