Combined protein for mycobacterium tuberculosis detection and detection reagent

A technology of Mycobacterium tuberculosis and combined protein, which is applied in the field of medical immune detection, can solve the problems of difficult promotion, high price, long detection period and the like

Active Publication Date: 2021-04-13
中国人民解放军总医院第八医学中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Based on this, the traditional separation and culture method often requires a long detection period, which delays clinical diagnosis and treatment, while the test results of γ-interferon release test need to be further excluded, Xpert-MTB / RIF test is expensive and difficult to popularize technical problems , the present invention proposes a combination protein and detection reagent for the detection of Mycobacterium tuberculosis

Method used

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  • Combined protein for mycobacterium tuberculosis detection and detection reagent
  • Combined protein for mycobacterium tuberculosis detection and detection reagent

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Embodiment 1

[0039]The preparation method of detection reagent for tuberculosis mycobacterium is as follows:

[0040]1), the synthetic gene fragment of the first combined protein of the present invention and the second composite protein of the present invention is cut from the cloned carrier, and XhoI is cut, cloned to the expression vector PET28B, in E. coli BL21 (DE3) The expression was incorporated to the OD600 was 0.6, and IPTG was added to the final concentration of 0.1 M induced expression, and analyzed by gel electrophoresis and WesternBlotting;

[0041]2), use gel (Pharmacia CHELANG SEPHROSE FAST FLOW), and the expression protein is purified by metallic chromatography;

[0042]3), using standard immunization procedural immunized rabbits, preparation of rabbits (IgM, IGA) by standard immunization programs (purchased from Tiangen Biocuro).

[0043]4), prepared by citric acid reducing method; ie, 0.01% aqueous solution of aqueous solution of aqueous solution of aqueous solution is heated, and 1% aqueou...

Embodiment 2

[0053]The preparation method of detection reagent for tuberculosis mycobacterium is as follows:

[0054]1), the synthetic gene fragment of the first combined protein of the present invention and the second composite protein of the present invention is cut from the cloned carrier, and XhoI is cut, cloned to the expression vector PET28B, in E. coli BL21 (DE3) The expression was incorporated to the OD600 was 0.6, and IPTG was added to the final concentration of 0.1 M induced expression, and analyzed by gel electrophoresis and WesternBlotting;

[0055]2), use gel (Pharmacia CHELANG SEPHROSE FAST FLOW), and the expression protein is purified by metallic chromatography;

[0056]3), using standard immunization procedural immunized rabbits, preparation of rabbits (IgM, IGA) by standard immunization programs (purchased from Tiangen Biocuro).

[0057]4), prepared by citric acid reducing method; ie, 0.01% aqueous solution of aqueous solution of aqueous solution of aqueous solution is heated, and 1% aqueou...

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Abstract

The invention provides combined protein for mycobacterium tuberculosis detection and a detection reagent. The combined protein is combined protein formed by combining TBCG-03312 protein or part of amino acids of the TBCG-03312 protein with any two or more of a B cell antigenic determinant of Rv1681 antigen protein, a B cell antigenic determinant of Rv1932 antigen protein, a B cell antigenic determinant of Rv1956 antigen protein and a B cell antigenic determinant of Rv3368c antigen protein through soft linkers. The formed combined protein has higher sensitivity and specificity; the combined protein is applied to a rapid detection method, the serum of suspected abortive tuberculosis patients can be more accurately and more rapidly detected so as to rapidly and accurately assist in diagnosis and treatment of the abortive tuberculosis patients, so that the cure rate of the tuberculosis patients is increased, and propagation of tuberculosis is avoided.

Description

Technical field[0001]The present invention belongs to the field of medical immunoassay, and more particularly to a combination protein and detection reagent for detecting mycobacterium tuberculosis.Background technique[0002]Despite 90 years of vaccination and history of chemotherapy, tuberculosis remains the main reason for death due to infection. Since 2007, it has been in the death of a single infectious disease. The World Health Organization estimates that the global junction latent infected population is about 1.7 billion, accounting for about 1 / 4 of the entire population. About 10.4 million new cases in 2018, 1.7 million people died in tuberculosis. One-third of these new cases (about 3 million people) is still not known for the health system, and many people have not obtained appropriate treatment.[0003]As long as the diagnosis and correct treatment is timely, almost all patients with drug sensitive tuberculosis can be cured, but the time to diagnose the diagnosis of the diagn...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/35C12N15/31G01N33/569G01N33/543
CPCC07K14/35G01N33/5695G01N33/54346G01N2333/35
Inventor 张灵霞孙卫国李夏南熊志红朱琰黄华董恩军郝彦斐
Owner 中国人民解放军总医院第八医学中心
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