Antigen polypeptide pool for detecting Mycobacterium tuberculosis infection

A technology of mycobacterium tuberculosis and antigenic polypeptides, applied in the biological field, can solve the problems of increasing detection sensitivity and insufficient detection rate, and achieve the effects of convenient operation, high detection rate and high sensitivity

Active Publication Date: 2017-09-29
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

[0009] 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 an

Method used

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  • Antigen polypeptide pool for detecting Mycobacterium tuberculosis infection
  • Antigen polypeptide pool for detecting Mycobacterium tuberculosis infection
  • Antigen polypeptide pool for detecting Mycobacterium tuberculosis infection

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0032] Example 1 Single polypeptide stimulation effectiveness screening

[0033] The amino acid sequence of the polypeptide sequence of the present invention is as shown in SEQ ID NO. 1 to SEQ ID NO. 26. These can be artificially synthesized or isolated naturally. The polypeptides used in this example and the following examples are artificially synthesized of.

[0034] SEQ ID NO.1: RLWVYCGNGTPNEL

[0035] SEQ ID NO.2: LMIGTAAAVVLPGL

[0036] SEQ ID NO.3: VQFQSGGNNSPAVY

[0037] SEQ ID NO.4: MPVGGQSSFYSDWY

[0038] SEQ ID NO.5: SAAIGLSMAGSSAM

[0039] SEQ ID NO.6: PAFEWYYQSGLSIV

[0040] SEQ ID NO.7: YQSAIPPRGTQAVV

[0041] SEQ ID NO.8: IQMSDPAYNINISL

[0042] SEQ ID NO.9: KSLENYIAQTRDKF

[0043] SEQ ID NO.10: MLVTAVVLLCCSGV

[0044] SEQ ID NO.11: RKAFRRAITRPTHF

[0045] SEQ ID NO.12: MVDPQLDGPQLAAL

[0046] SEQ ID NO.13: KLDSPIIARITDTV

[0047] SEQ ID NO.14: RLAAQTALLESEAL

[0048] SEQ ID NO.15: ITIAVNADSMATWF

[0049] SEQ ID NO.16: REKPCRATTAGIPL

[0050] SEQ ID NO.17: LLFPAIYLADSAQA

[0051] SEQ I...

Example Embodiment

[0063] Example 2 Combined stimulation effect

[0064] Combine any 8 polypeptide solutions with ESAT-6 and CFP-10 to prepare a stimulant solution, where ESAT-6 and CFP-10 are obtained by recombinant expression of E. coli genetically engineered bacteria, and the ESAT-6 sequence is SEQ ID NO. 27 :MTEQQWNFAGIEAAASAIQGNVTSIHSLLDEGKQSLTKLAAAWGGSGSEAYQGVQQKWDATATELNNALQNLARTISEAGQAMASTEGNVTGMFA; The sequence of CFP-10 is SEQ ID NO.28: MAEMKTDAATLAQEAGNISGDLKTQIDFERQVESTAGSLQGQWRGAQELQEAQGRFEEQRFEEQFREEQGRFKG In this example, only part of the experimental results of polypeptides are listed, numbered as stimulant 1 to stimulant 4 (stimulant 1: SEQ ID NO. 1 to SEQ ID NO. 8+ESAT-6+CFP-10; stimulant 2: SEQ ID NO.6~SEQID NO.13+ESAT-6+CFP-10; Stimulant 3: SEQ ID NO.11~SEQ ID NO.18+ESAT-6+CFP-10; Stimulant 4: SEQ ID NO .19~SEQ ID NO.26+ESAT-6+CFP-10), take positive and uninfected negative samples of Mycobacterium tuberculosis infection for testing.

[0065] Take the 20 whole blood samples from ...

Example Embodiment

[0069] Example 3 Combined stimulation effect

[0070] Combine any eight polypeptide solutions with ESAT-6-CFP-10 to prepare a stimulant solution, where ESAT-6-CFP-10 is obtained by recombinant expression of E. coli genetically engineered bacteria, and the sequence of ESAT-6-CFP-10 is SEQ ID NO.29: MTEQQWNFAGIEAAASAIQGNVTSIHSLLDEGKQSLTKLAAAWGGSGSEAYQGVQQKWDATATELNNALQNLARTISEAGQAMASTEGNVTGMFANVAMAEMKTDAATLAQEAGNFERISGDLKTQIDQVESTAGSLQGQWRGAAGTAAQAAVVRFQEAANKQKQELDEISTNIRQAGVQYSRADEEQQQALSSQMGF, stimulants No. 5 to 8 stimulators (stimulator 5: SEQ ID NO.1 ~ SEQ ID NO.8 + ESAT-6-CFP-10; stimulators 6: SEQ ID NO.6 ~SEQ ID NO.13+ESAT-6-CFP-10; Stimulant 7: SEQ ID NO.11~SEQ ID NO.18+ESAT-6-CFP-10; Stimulant 8: SEQ IDNO.19~SEQ ID NO.26+ESAT-6-CFP-10), take the positive and negative samples of Mycobacterium tuberculosis for testing, prepare the stimulant solution, take the 20 whole blood samples as in Example 1, and use the gamma interferon detection reagent The box detects whether it i...

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Abstract

The invention discloses an antigen polypeptide pool for detecting Mycobacterium tuberculosis infection. The specific antigen polypeptide pool can specifically stimulate Mycobacterium-tuberculosis-infected fresh whole blood and specifically secrete IFN-gamma, thereby increasing the detection sensitivity. The invention provides a novel detection reagent applicable to detection of Mycobacterium tuberculosis infection. The experiment proves that the peripheral blood can be directly utilized to perform antigenic stimulation without the need for separating peripheral blood mononuclear cells. The experimental data shows that when being used for detecting Mycobacterium tuberculosis infection, the antigen polypeptide pool has the advantages of higher sensitivity, higher specificity and lower cost, is simple to operate, and has higher 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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IPC IPC(8): C07K14/35G01N33/569G01N33/68
CPCC07K14/35G01N33/5695G01N33/6866G01N2333/35
Inventor 张诗冉丁晓莉路春桃赵平锋
Owner 武汉海吉力生物科技有限公司
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