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Antigenic polypeptide pool for detecting mycobacterium tuberculosis infection and application

A technology of mycobacterium tuberculosis and antigenic polypeptides, applied in the biological field, can solve the problems of stimulation, insufficient detection rate, increase detection sensitivity, etc., and achieve the effect of rapid diagnosis, convenient operation and high sensitivity

Active Publication Date: 2017-09-08
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 sex antigen peptide pool specifically stimulates the fresh whole blood infected with Mycobacterium tuberculosis to specifically secrete IFN-γ, increasing the detection sensitivity

Method used

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  • Antigenic polypeptide pool for detecting mycobacterium tuberculosis infection and application
  • Antigenic polypeptide pool for detecting mycobacterium tuberculosis infection and application
  • Antigenic polypeptide pool for detecting mycobacterium tuberculosis infection and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Screening of Single Polypeptide Stimulatory Effectiveness

[0032] The amino acid sequence of the polypeptide of the present invention is shown in SEQ ID NO.1 to SEQ ID NO.28. These can be artificially synthesized or naturally isolated. The polypeptides used in this example and the following examples are artificially synthesized .

[0033] SEQ ID NO. 1: ALGSSLHTAGVDLA

[0034] SEQ ID NO. 2: RLGVLASHHDNAAV

[0035] SEQ ID NO. 3: NVYLTAHNALGSSL

[0036] SEQ ID NO. 4: ITHGPYCSQFNDTL

[0037] SEQ ID NO.5: SHHDNAAVDASSGV

[0038] SEQ ID NO. 6: RIAAKIYSEADEAWR

[0039] SEQ ID NO. 7: NALGSSLHTAGVDLA

[0040] SEQ ID NO. 8: ALAAVVELGSFDAA

[0041] SEQ ID NO.9: LMAGAGPAPMLAAA

[0042] SEQ ID NO. 10: AALDAQAVELTARL

[0043] SEQ ID NO. 11: LMSQLIEKPVAPSV

[0044] SEQ ID NO. 12: SLPEIAANHITQAV

[0045] SEQ ID NO. 13: MQATAQAAAYTQAM

[0046] SEQ ID NO. 14: KLAGLVFPQPPAPI

[0047] SEQ ID NO. 15: QQAAQSAQGGSGPM

[0048] SEQ ID NO. 16: VQMSQNASPIAQTI

[0049] SE...

Embodiment 2

[0064] Embodiment 2 merges stimulation effect

[0065] Combine ESAT-6 and CFP-10 with any 8 polypeptide solutions to prepare a stimulator solution (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. The sequence of CFP-10 is SEQ ID NO.30: MAEMKTDAATLAQEAGNFERISGDLKTQIDQVESTAGSLQGQWRGAAGTAAQAAVVRFQEAANKQKQELDEISTNIRQAGVQYSRADEEQQQALSSQMGF. In this embodiment, only the experimental results of some polypeptides are listed, numbered as stimulator 1 to stimulator 4 (stimulator 1: SEQ ID NO.1 to SEQ ID NO. 8+ESAT-6+CFP-10; Stimulator 2: SEQ ID NO.8~SEQ ID NO.15+ESAT-6+CFP-10; Stimulator 3: SEQ ID NO.13~SEQ ID NO.20+ESAT -6+CFP-10; stimulant 4: SEQ ID NO.21~SEQ ID NO.28+ESAT-6+CFP-10;), take the positive and non-infected negative samples of Mycobacterium tuberculosis infection for testing.

[0066] Take the same 20 cases of whole blood samples in Example 1, and use the gamma interfero...

Embodiment 3

[0070] Embodiment 3 merges stimulation effect

[0071]将任取8条多肽溶液合并ESAT-6-CFP-10(ESAT-6-CFP-10由大肠杆菌基因工程菌重组表达获得,ESAT-6-CFP-10序列为SEQ ID NO.31:MTEQQWNFAGIEAAASAIQGNVTSIHSLLDEGKQSLTKLAAAWGGSGSEAYQGVQQKWDATATELNNALQNLARTISEAGQAMASTEGNVTGMFANVAMAEMKTDAATLAQEAGNFERISGDLKTQIDQVESTAGSLQGQWRGAAGTAAQAAVVRFQEAANKQKQELDEISTNIRQAGVQYSRADEEQQQALSSQMGF, Prepare the stimulating agent solution, numbered as stimulating agent 5~stimulating agent 8 (stimulating agent 5: SEQ ID NO.1~SEQ ID NO.8+ESAT-6-CFP-10; stimulating agent 6: SEQ ID NO.8~SEQ ID NO. ID NO.15+ESAT-6-CFP-10; Stimulator 7: SEQ ID NO.13~SEQ ID NO.20+ESAT-6-CFP-10; Stimulator 8: SEQ ID NO.21~SEQ ID NO .28+ESAT-6-CFP-10), get the positive and uninfected negative samples of mycobacterium tuberculosis infection to test, be mixed with stimulant solution, get with 20 routine whole blood samples in embodiment 1, use γ Interferon detection kit detects whether it infects Mycobacterium tuberculosis. At the same time, take 20 μL each of stimula...

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Abstract

The invention discloses an antigenic polypeptide pool for detecting mycobacterium tuberculosis infection. The specific antigenic polypeptide pool can specifically secrete IFN-gamma by specifically stimulates fresh whole blood infected by the mycobacterium tuberculosis to improve the detection sensitivity. The invention provides a new detection kit applied to detection of mycobacterium tuberculosis infection. Experiments prove that antigenic stimulation can be conducted by directly using peripheral blood, single karyocyte of the peripheral blood does not need to be separated, experiment data shows that the antigenic polypeptide pool has high sensitivity and specificity when being used for detecting mycobacterium tuberculosis infection, is simple and convenient to operate, and low in cost, and has high clinical application value.

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
CPCC07K14/35G01N33/5695G01N33/6866G01N2333/57
Inventor 丁晓莉路春桃张诗冉熊俊赵平锋
Owner 武汉海吉力生物科技有限公司
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