Application of brucellosis A19 molecular marking vaccine and immunological identification thereof

A technique for molecularly labeling vaccines and brucellosis, applied to bacterial antigen components, analytical materials, antibacterial drugs, etc., can solve the problem of difficult identification of immunized animals and clinically diseased animals

Inactive Publication Date: 2012-11-14
新疆维吾尔自治区畜牧科学院兽医研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The brucellosis A19-ΔVirB12 labeled vaccine provided by the invention not only has good immune protection against bovine brucellosis, but also the iELISA method invented solves the problem that it is difficult to distinguish between immunized animals and clinically ill animals, and is effective for bovine brucellosis. The prevention, control, eradication and purification of brucellosis have practical application value

Method used

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  • Application of brucellosis A19 molecular marking vaccine and immunological identification thereof
  • Application of brucellosis A19 molecular marking vaccine and immunological identification thereof
  • Application of brucellosis A19 molecular marking vaccine and immunological identification thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The application of brucellosis A19 molecular marker vaccine and its immune protection:

[0028] Material:

[0029] Strains: A19-ΔVirB12 labeled vaccine was developed and preserved by the Veterinary Research Institute of Xinjiang Academy of Animal Sciences, Brucella 2308 was purchased from China Veterinary Drug Control Institute, and A19 vaccine was purchased from Xinjiang Tiankang Animal Husbandry Biotechnology Co., Ltd.;

[0030] Experimental animals: 25 Xinjiang native cattle aged 6-8 months, purchased from the live cattle trading market in Miquan District, Xinjiang, all tested negative for brucellosis;

[0031] Determination of the challenge dose of Brucella 2308 strain: 25 Xinjiang local cattle were randomly divided into 5 groups, 5 cattle in each group, among which, the 1-4 groups were challenged with Brucella 2308 standard strain, eyelid The inoculated challenge dose was 1×10 6 CFU, 1×10 7 CFU, 1×10 8 CFU, 1×10 9 CFU, the fifth group is the blank control grou...

Embodiment 2

[0042] Immunological identification of brucellosis A19 molecular marker vaccine:

[0043] Preparation of VirB12 antigen protein:

[0044] Strains and vectors: A19 molecularly labeled vaccine was developed by the Veterinary Research Institute of Xinjiang Academy of Animal Sciences; Brucella 2308 strains were purchased from the China Veterinary Drug Administration; the cloned pGEM-T vector was purchased from Promega as a commercial reagent; prokaryotic expression vectors pET-28a(+) is preserved by the Veterinary Research Institute of Xinjiang Academy of Animal Husbandry Science;

[0045] Reagents: Restriction enzymes were purchased from Fermentas; plasmid DNA extraction kit and DNA gel recovery kit were purchased from Promega; T4DNA ligase, DNA Marker, and DH5α were products of Beijing Dingguo Biotechnology Company. The enzyme-labeled secondary antibody was rabbit anti-bovine IgG horseradish peroxidase (Bethyl, USA); the His-trap column for protein purification was a product of...

Embodiment 3

[0054] iELISA identification of A19-ΔVirB12 vaccine antibody and field strain antibody:

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Abstract

The invention relates to an application of brucellosis A19 molecular marking vaccine and immunological identification of the brucellosis A19 molecular marking vaccine. The brucellosis vaccine can be applied to an enzyme-linked immunosordent assay (iELISA) method to distinguish an animal inoculated with the vaccine from a wild virus infected animal. According to the invention, a cattle is used as a test target animal antiepidemic to the brucellosis, the cattle is immune to brucellosis A19-detlaVirB12 marking vaccine so as to evaluate the immunological protection of the molecular marking vaccine. An iELISA method for distinguishing the brucellosis A19-detlaVirB12 immune animal from the wild virus infected animal is created. The brucellosis A19-detlaVirB12 marking vaccine provided by the invention not only has good immunological protection against the cattle brucellosis, and the iELISA method provided by the invention solves the problem that the immune animal is difficult to be distinguished from the clinic diseased animal, and the brucellosis A19 molecular marking vaccine has an actual application value in prevention and control, elimination and purification of the cattle brucellosis.

Description

technical field [0001] The present invention relates to the application of brucellosis A19 molecularly marked vaccine and its immunological identification, especially the (A19-ΔVirB12) immune protection and application of brucellosis A19 molecularly marked vaccine. The problem that it is difficult to distinguish between animals and clinically diseased animals belongs to the field of veterinary biological products. Background technique [0002] Brucellosis (Brucellosis) is caused by Brucella (Brucella) widespread zoonotic infectious diseases around the world. The prevention and eradication of brucellosis has become the main task of public health security defense systems in many countries and regions. [0003] In most countries and regions where brucellosis is endemic, the main means of controlling livestock brucellosis is still to use vaccination for immunization. Therefore, vaccination is the main measure and the most effective means to prevent and control livestock brucell...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/10A61P31/04G01N33/53G01N33/531G01N33/543C12N15/70C12R1/19C12R1/01
Inventor 钟旗易新萍王力俭谷文喜叶锋刘丽娅李延涛吴冬玲马晓菁吐尔洪·努尔姚刚范伟兴
Owner 新疆维吾尔自治区畜牧科学院兽医研究所
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