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

A technology for infecting cells and immune detection, applied in the field of tuberculosis detection, can solve the problems such as the need to improve the sensitivity and specificity, high cost, complex composition of antigen components, etc., to achieve improved detection sensitivity, low cost, and suitable for large-scale commercial production. Effect

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The composition of embodiment 1 epitope peptide composition

[0045] The composition for tuberculosis detection consists of epitope peptide derived from CFP-10 antigen, ESAT-6 antigen and Rv3873 antigen.

[0046] The amino acid sequence of the CFP-10 antigen is shown in SEQ ID NO.1.

[0047] The amino acid sequence of the ESAT-6 antigen is shown in SEQ ID NO.2.

[0048] The amino acid sequence of the Rv3873 antigen is shown in SEQ ID NO.3.

[0049] The amino acid sequence of the epitope peptide derived from the CFP-10 antigen is selected from SEQ ID NO.4-59.

[0050] The amino acid sequence of the epitope peptide derived from ESAT-6 antigen is selected from SEQ ID NO.60-122.

[0051] The amino acid sequence of the epitope peptide derived from the Rv3873 antigen is selected from SEQ ID NO.123-180.

Embodiment 2

[0052] Example 2 Antigen epitopes are applied to the development of detection kits

[0053] 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.

[0054] The test kit includes:

[0055] 1) the epitope peptide composition described in Example 1;

[0056] 2) mouse IgG monoclonal antibody (primary antibody) against human or animal IFN-γ, which is fixed on the reaction plate;

[0057] 3) Enzyme-labeled reagent: another mouse IgG monoclonal antibody (secondary antibody) labeled with horseradish peroxidase against different epitopes of human or animal IFN-γ;

[0058] 4) Calibrator: positive raw material containing IFN-γ;

[0059] 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 antigens, and the backg...

Embodiment 3

[0061] Example 3 Antigen epitopes are applied to clinical detection of tuberculosis infection

[0062] A. Isolation of peripheral blood lymphocytes:

[0063] The screening criteria of volunteer cases involved in the present invention are:

[0064] 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.) patients.

[0...

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Abstract

The invention provides an antigen composition used for immunodetection of a tuberculosis infected cell. The antigen composition comprises mycobacterium tuberculosis CFP-10, ESAT-6 and Rv3873 antigens and / or antigen epitope peptides derived from the antigens. In an optimized embodiment, the antigen composition comprises all the antigen epitope peptides contained in the mycobacterium tuberculosis CFP-10, ESAT-6 and Rv3873 antigens. 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
CPCA61K39/00C07K14/35G01N33/577G01N33/68
Inventor 万康林季绍有李马超蒋毅刘海灿
Owner ICDC CHINA CDC
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