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Protein capable of specifically detecting mycobacterium tuberculosis infection

A Mycobacterium tuberculosis protein technology, applied in the field of biomedicine, can solve the problems of insensitivity of latent infection diagnosis of Mycobacterium tuberculosis, unsatisfactory sensitivity and specificity of Mycobacterium tuberculosis antibody, etc.

Inactive Publication Date: 2017-01-25
TB HEALTHCARE BIOTECHNOLOGY (GUANGDONG) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the diagnosis of tuberculosis commonly used in clinical practice mainly relies on clinical symptoms, impact diagnosis and etiology diagnosis, and is not sensitive to the diagnosis of latent infection of Mycobacterium tuberculosis.
At the same time, in the process of TB screening, the sensitivity and specificity of direct detection of pathogens or detection of Mycobacterium tuberculosis antibodies are not ideal

Method used

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  • Protein capable of specifically detecting mycobacterium tuberculosis infection
  • Protein capable of specifically detecting mycobacterium tuberculosis infection
  • Protein capable of specifically detecting mycobacterium tuberculosis infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1, prokaryotic expression of target protein RV1438

[0036] Preparation of the target protein RV1438: Introduce the recombinant vector pET28a-RV1438 expressing the gene encoding the RV1438 protein into Escherichia coli BL21 to obtain the recombinant strain BL21 / pET28a-RV1438; then induce the recombinant strain BL21 / pET28a-RV1438 with IPTG, and collect the supernatant, namely For the target protein RV1438.

[0037] details as follows:

[0038] 1. Construction of recombinant bacteria expressing the target protein

[0039] The amino acid sequence of the RV1438 protein is sequence 2, and the nucleotide sequence of its coding gene is sequence 1.

[0040] Insert the RV1438 protein coding gene shown in Sequence 1 into the NdeI and EcoRI sites of the pET28a vector to obtain the recombinant vector pET28a-RV1438;

[0041] The recombinant vector was introduced into Escherichia coli BL21 to obtain recombinant bacterium BL21 / pET28a-RV1438.

[0042] 2. Protein expressi...

Embodiment 2

[0061] Example 2, the application of Mycobacterium tuberculosis marker antigen RV1438 in the detection of Mycobacterium tuberculosis

[0062] 1. T-SPOT kit detection

[0063] details as follows:

[0064] details as follows:

[0065] 1) Use heparin vacuum blood collection tubes to directly collect peripheral venous blood from different patients (as shown in Table 1, and the patients are informed) to obtain anticoagulant blood samples;

[0066] 2) Mix the anticoagulant blood sample with RPMI1640 culture medium at a volume ratio of 1:1; carefully add the blood sample to the upper layer of Ficoll lymphocyte separation medium at a ratio of 3:1 (using Ficoll-PaqueTM plus Separation of PBMCs according to the operating instructions); room temperature 25 ° C, 1000g centrifugation for 22 minutes; horizontal rotor, slow up and down;

[0067] 3) Transfer the mononuclear cell layer (located in the middle layer of the centrifuge tube, in the shape of a thin white film) from the Ficoll se...

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Abstract

The invention discloses protein capable of specifically detecting mycobacterium tuberculosis infection, and provides application of Rv1438 protein to preparation of products for detecting or assisting in detecting mycobacterium tuberculosis. The experiment of the invention proves that mycobacterium tuberculosis-derived protein segments are screened, a mycobacterium tuberculosis marker antigen RV1438 for detecting tuberculosis is provided, and the antigen is used for in vitro detecting specific T cell immunoreaction to serve as a reference for diagnosing patients suffering from tuberculosis and can be used for diagnosing whether the patients are infected with the mycobacterium tuberculosis. The protein can be used for detecting mycobacterium tuberculosis infection, and the specificity reaches 70% at present.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to a protein for specifically detecting mycobacterium tuberculosis infection. Background technique [0002] my country's in vitro diagnostic industry is now in a period of rapid growth. However, there is an obvious gap between the huge market demand and the relatively backward independent research and development capabilities of my country's diagnostic reagent industry. How to carry out independent innovation from the source, how to not be restricted by individual overseas pharmaceutical giants in the supply of raw materials for diagnostic reagents, and how to transform existing scientific research achievements in the field of biomedicine into products have become the key issues for the Chinese government, scientific research institutes and the pharmaceutical industry in recent years. hot spots of great concern. Moreover, there is a rigid demand for more sensitive and efficient diagnost...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/35C12N15/31G01N33/569G01N33/50G01N33/68A61K39/04A61P31/06
CPCC07K14/35A61K39/00G01N33/505G01N33/5695G01N33/68G01N2800/12
Inventor 毕利军张先恩朱国峰邓教宇陶生策侯剑王雅果
Owner TB HEALTHCARE BIOTECHNOLOGY (GUANGDONG) CO LTD
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