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Antigenic polypeptide pool capable of detecting mycobacterium tuberculosis infection, and application thereof

A technology of Mycobacterium tuberculosis and antigenic polypeptides, applied in the biological field, can solve problems such as stimulation, insufficient detection rate, and increase detection sensitivity, and achieve the effects of rapid diagnosis, convenient operation and high sensitivity

Active Publication Date: 2017-08-04
武汉海吉力生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In view of the insufficient detection rate of existing detection methods for some special samples, such as children, immunocompromised and newly infected samples of Mycobacterium tuberculosis, the purpose of the present invention is to provide an antigen polypeptide pool for detecting Mycobacterium tuberculosis infection, the antigen The peptide pool specifically stimulates the whole blood infected with Mycobacterium tuberculosis to specifically secrete IFN-γ, increasing the detection sensitivity

Method used

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  • Antigenic polypeptide pool capable of detecting mycobacterium tuberculosis infection, and application thereof
  • Antigenic polypeptide pool capable of detecting mycobacterium tuberculosis infection, and application thereof
  • Antigenic polypeptide pool capable of detecting mycobacterium tuberculosis infection, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Screening of Single Polypeptide Stimulatory Effectiveness

[0032] The polypeptide of the present invention has an amino acid sequence as shown in SEQ ID NO.1 to SEQ ID NO.29, which can be artificially synthesized or naturally isolated. The polypeptides used in this example and the following examples are artificially synthesized , wherein, SEQ ID NO.1~SEQ ID NO.5 is from Ag85B, SEQ ID NO.6 is from Mpt649, SEQ ID NO.7~SEQ ID NO.8 is from RV1985C, SEQ ID NO.9~SEQ ID NO.10 is from RV1989c, SEQ ID NO.11~SEQ ID NO.12 are from Rv3425, SEQ ID NO.13~SEQ ID NO.15 are from RV3615, SEQ ID NO.16~SEQ NO.19 are from RV3873, SEQ ID NO.20~SEQ ID NO.22 is from RV3878, SEQ ID NO.23 is from RV3879c, SEQ ID NO.24~SEQ ID NO.26 is from tb7.7, SEQ ID NO.27 is from CFP-10, SEQ ID NO.28~SEQ IDNO.29 From ESAT-6.

[0033] SEQ ID NO.1: RLWVYCGNGTPNEL

[0034] SEQ ID NO. 2: LMIGTAAAVVLPGL

[0035] SEQ ID NO. 3: VQFQSGGNNSPAVY

[0036] SEQ ID NO. 4: MPVGGQSSFYSDWY

[0037] SEQ ID NO....

Embodiment 2

[0065] Embodiment 2 merges stimulation effect

[0066] Combine ESAT-6 and CFP-10 with any 8 polypeptide solutions (ESAT-6 and CFP-10 are obtained by recombinant expression of Escherichia coli genetically engineered bacteria, and the sequence of ESAT-6 is SEQ ID NO. It is SEQ ID NO.31: MAEMKTDAATLAQEAGNFERISGDLKTQIDQVESTAGSLQGQWRGAAGTAAQAAVVRFQEAANKQKQELDEISTNIRQAGVQYSRADEEQQQALSSQMGF, which is prepared as a stimulating agent solution. In this embodiment, only the experimental results of preferred partial polypeptides are listed, numbered as stimulating agent 1~stimulating agent 4 (stimulating agent 1: SEQ ID NO.1~SEQ ID NO.8+ESAT-6+CFP-10; Stimulator 2: SEQ ID NO.9~SEQ ID NO.16+ESAT-6+CFP-10; Stimulant 3: SEQ ID NO.17~SEQ ID NO. 24+ESAT-6+CFP-10; Stimulator 4: SEQ ID NO.22~SEQ ID NO.29+ESAT-6+CFP-10), the positive and uninfected negative samples of Mycobacterium tuberculosis infection were taken for test.

[0067] Take the same 20 cases of whole blood samples in Example 1, a...

Embodiment 3

[0071] Embodiment 3 merges stimulation effect

[0072] Combine any 8 polypeptide solutions with ESAT-6-CFP-10 to prepare a stimulator solution, wherein ESAT-6-CFP-10 is obtained by recombinant expression of Escherichia coli genetically engineered bacteria, and the sequence of ESAT-6-CFP-10 is SEQ ID NO.32:

[0073]MTEQQWNFAGIEAAASAIQGNVTSIHSLLDEGKQSLTKLAAAWGGSGSEAYQGVQQKWDATATELNNALQNLARTISEAGQAMASTEGNVTGMFANVAMAEMKTDAATLAQEAGNFERISGDLKTQIDQVESTAGSLQGQWRGAAGTAAQAAVVRFQEAANKQKQELDEISTNIRQAGVQYSRADEEQQQALSSQMGF,编号为刺激剂5~刺激剂8(刺激剂5:SEQ ID NO.1~SEQ ID NO.8+ESAT-6-CFP-10;刺激剂6:SEQ ID NO.9~SEQ IDNO.16 +ESAT-6-CFP-10; Stimulator 7: SEQ ID NO.17~SEQ ID NO.24+ESAT-6-CFP-10; Stimulator 8: SEQ ID NO.22~SEQ ID NO.29+ESAT -6-CFP-10), get the positive and non-infected negative samples of mycobacterium tuberculosis infection to test, be mixed with stimulant solution, get with 20 routine whole blood samples among the embodiment 1, use gamma interferon detection reagent The box detects whether ...

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Abstract

The invention discloses an antigenic polypeptide pool capable of detecting mycobacterium tuberculosis infection. The antigenic polypeptide pool specifically stimulates mycobacterium tuberculosis infected fresh whole blood to specifically secrete IFN-gama and increase the sensitivity. The invention provides a new detection reagent for detecting mycobacterium tuberculosis infection, experiments prove that peripheral blood can be directly utilized to conduct antigenic simulation without separation of peripheral blood mononuclear cell, the experimental data shows that the antigenic polypeptide pool has high sensitivity and specificity for detecting mycobacterium tuberculosis infection, and is simple and convenient to operate and low in cost, thus having high clinical application values.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an antigen polypeptide pool for detecting mycobacterium tuberculosis infection and its application. Background technique [0002] Tuberculosis is one of the important infectious diseases that are mainly transmitted through the respiratory tract and seriously endanger my country and the world. According to the World Health Organization, there are 1.7-2 billion Mycobacterium tuberculosis infections in the world, and at least 2 million people die from this disease every year (Xie Jianping, Methodology for Mycobacterium tuberculosis functional genome research, Microbiology Bulletin. 2001.28 (5) :92-97). Therefore, methods of accurately screening infected persons play an extremely important role in the prevention and control and even the eventual elimination of tuberculosis. [0003] Existing diagnostic techniques are very difficult to diagnose patients with Mycobacterium tuberculosis i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/35G01N33/569G01N33/68
CPCC07K14/35G01N33/5695G01N33/6866G01N2333/35
Inventor 黄俊陈银芳张诗冉丁晓莉赵平锋
Owner 武汉海吉力生物科技有限公司
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