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Antigen used for immunodetection of tuberculosis infected T cell and application thereof

A technology for infecting cells and immunological detection, applied in the field of tuberculosis detection, can solve the problems of high cost, need to improve sensitivity and specificity, and complex composition of antigen components, and achieve low cost, improved detection sensitivity, and true and reliable detection results

Active Publication Date: 2015-05-20
ICDC CHINA CDC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Based on the in vitro detection technology of interferon gamma secreted by T cells, the intact antigen is mainly used as the corresponding stimulus source, the composition of the antigen is relatively complex, and the sensitivity and specificity need to be improved, and the corresponding reagents are patented by foreign companies. The cost of product application is relatively high, and it is not suitable for large-scale application in my country under the current situation

Method used

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  • Antigen used for immunodetection of tuberculosis infected T cell and application thereof
  • Antigen used for immunodetection of tuberculosis infected T cell and application thereof
  • Antigen used for immunodetection of tuberculosis infected T cell and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0031] The composition of embodiment 1 epitope peptide composition

[0032] Antigen epitope peptide composition for tuberculosis detection consists of epitope peptide derived from Rv3873 antigen. The amino acid sequence of the antigen Rv3873 is shown in SEQ ID NO: 1; the amino acid sequence of the epitope peptide derived from the Rv3873 antigen is shown in SEQ ID NO: 2-59.

Embodiment 2

[0033] Example 2 Antigen epitope peptide composition is applied to the development of detection kit

[0034] The antigen epitope peptide composition of the present invention can be used for the detection of tuberculosis infection, and can form a kit for clinical or laboratory diagnosis.

[0035] The test kit includes:

[0036]1) the epitope peptide composition described in Example 1;

[0037] 2) mouse IgG monoclonal antibody (primary antibody) against human or animal IFN-γ, which is fixed on the reaction plate;

[0038] 3) Enzyme-labeled reagent: another mouse IgG monoclonal antibody (secondary antibody) labeled with horseradish peroxidase against different epitopes of human or animal IFN-γ;

[0039] 4) Calibrator: positive raw material containing IFN-γ;

[0040] 5) Culture tubes: the test culture tubes contain artificially synthesized epitope polypeptides derived from various antigens, the positive control culture tubes contain tuberculosis non-specific stimulating antigen...

Embodiment 3

[0042] Example 3 Application of Antigenic Epitope Polypeptide in Clinical Detection of Tuberculosis Infection

[0043] A. Isolation of peripheral blood lymphocytes:

[0044] The screening criteria of volunteer cases involved in the present invention are:

[0045] According to the national standard (WS288-2008) of the "Diagnostic Criteria for Pulmonary Tuberculosis" issued by the Ministry of Health, patients with clinical manifestations, symptoms, signs and chest imaging examinations diagnosed as pulmonary tuberculosis, among which bacteriologically positive or negative tuberculosis patients refer to patients clinically diagnosed as tuberculosis Among them, tuberculosis patients with positive or negative bacteriological examination (microscopic examination of sputum smear acid-fast staining and / or bacterial solid culture on Roche solid medium); and extrapulmonary tuberculosis (such as bone tuberculosis, renal tuberculosis, intestinal tuberculosis, lymphatic tuberculosis) etc.)...

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Abstract

The invention provides an antigen composition used for immunodetection of a tuberculosis infected cell. The antigen composition comprises a mycobacterium tuberculosis Rv3873 antigen and / or antigen epitope peptides derived from the antigen. In an optimized embodiment, the antigen composition comprises all the antigen epitope peptides contained in the mycobacterium tuberculosis Rv3873 antigen. The invention also provides an application of the antigen composition to preparation of detection reagents used for detecting specific T cell immune response caused by tuberculosis infection in vitro. After lymphocytes of human or animals are stimulated by the antigen composition, cell factors secreted by a tuberculosis specific T cell are detected.

Description

technical field [0001] The invention relates to a specific T cell immune detection antigen for tuberculosis infection and its application. Specifically, it is a mycobacterium tuberculosis antigen and its epitope peptide composition, which is used for immune detection of tuberculosis infected cells and belongs to the field of tuberculosis detection. Background technique [0002] Tuberculosis is a zoonotic chronic infectious disease caused by Mycobacterium tuberculosis infection. According to the statistics of the World Health Organization, more than 1 / 3 of the world's population is currently infected with Mycobacterium tuberculosis, and 2 million people die from tuberculosis every year. my country is one of the 22 countries with severe tuberculosis epidemics in the world, and it is also one of the 27 countries with severe multidrug-resistant tuberculosis epidemics in the world. According to the fifth sampling survey report on tuberculosis epidemiology in China, the annual n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/35C12N15/31C12N15/63C12N1/21G01N33/68G01N33/577A61K39/04A61P31/04C12R1/32
CPCA61K39/00C07K14/35G01N33/5695G01N33/68G01N2333/35
Inventor 万康林李马超季绍有刘海灿蒋毅
Owner ICDC CHINA CDC
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