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Protein rv2818c for specific detection of Mycobacterium tuberculosis infection

A technology of Mycobacterium tuberculosis and rv2818c, which is applied in the field of biomedicine, can solve the problems that active pulmonary tuberculosis cannot be diagnosed and the positive detection rate is low.

Active Publication Date: 2022-02-11
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the antigens currently targeted for serological diagnosis of Mycobacterium tuberculosis, such as antigen 5, 38KD antigen, 30 / 31KD antigen, and antigen 60, all have problems such as low positive detection rate and cross-immunity with other mycobacteria. It has certain value in monitoring, prompting recurrence, judging prognosis and general screening of high-risk groups, but it still cannot be used for the diagnosis of active pulmonary tuberculosis

Method used

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  • Protein rv2818c for specific detection of Mycobacterium tuberculosis infection
  • Protein rv2818c for specific detection of Mycobacterium tuberculosis infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1, prokaryotic expression of target protein Rv2818c

[0039] Preparation of the target protein Rv2818c: Introduce the recombinant vector pET28a-Rv2818c expressing the Rv2818c protein coding gene into Escherichia coli BL21 to obtain the recombinant strain BL21 / pET28a-Rv2818c; then induce the recombinant strain BL21 / pET28a-Rv2818c with IPTG, and collect the supernatant, namely For the target protein Rv2818c.

[0040] details as follows:

[0041] 1. Construction of recombinant bacteria expressing the target protein

[0042] The amino acid sequence of the Rv2818c protein is sequence 2, and the nucleotide sequence of its coding gene is sequence 1.

[0043] Insert the Rv2818c protein coding gene shown in Sequence 1 into the NdeI and EcoRI sites of the pET28a vector to obtain the recombinant vector pET28a-Rv2818c;

[0044] The recombinant vector was introduced into Escherichia coli BL21 to obtain the recombinant bacterium BL21 / pET28a-Rv2818c.

[0045] 2. Protein ex...

Embodiment 2

[0060] Example 2, the application of Mycobacterium tuberculosis marker antigen Rv2818c in the detection of Mycobacterium tuberculosis

[0061] 1. T-SPOT kit detection

[0062] The specific method is as follows, and some solutions inside are all from the T-SPOT kit of Oxford immunotec (UK):

[0063] 1) Use heparin vacuum blood collection tubes to directly collect peripheral veins from different patients to be tested (61 patients with tuberculosis clinically detected as shown in Table 1 and 10 cases of clinically detected non-tuberculosis patients shown in Table 2, and informed). blood, to obtain anticoagulated blood samples;

[0064] 2) Mix the anticoagulated blood sample with RPMI1640 culture medium at a volume of 1:1; carefully add the blood sample to the upper layer of Ficoll lymphocyte separation medium (GE Healthcare 17-1440-02) at a ratio of 3:1 (using Ficoll-PaqueTM plus Separation of PBMCs according to the operating instructions); room temperature 25 ° C, 1000g centri...

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Abstract

The invention discloses protein Rv2818c for specifically detecting tuberculosis mycobacterium infection. The present invention provides the application of Rv2818c protein in the preparation, detection or auxiliary detection of Mycobacterium tuberculosis products; the Rv2818c is the following a) or b) protein: a) a protein composed of the amino acid sequence shown in sequence 1 in the sequence table ; b) a protein derived from a) that undergoes substitution and / or deletion and / or addition of one or several amino acid residues in the amino acid residue sequence of Sequence 1 in the sequence listing and has the same function as the protein shown in Sequence 1 . Experiments of the present invention prove that the detection rate of mycobacterium tuberculosis infection can be more effectively improved.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to protein Rv2818c for specifically detecting tuberculosis mycobacterium infection. Background technique [0002] For centuries, tuberculosis has continued to be a global public health problem that cannot be ignored. At present, one-third of the world's population is carrying Mycobacterium tuberculosis. In 2010 alone, there were 8.8 million new tuberculosis cases and 1.45 million deaths. On average, one person died of tuberculosis in less than 22 seconds. Tuberculosis is the highest infectious disease The number one death toll. [0003] Scientific evidence shows that due to the high transmissibility of active tuberculosis, it is of great significance to diagnose active tuberculosis, eliminate the source of infection as soon as possible, and control the epidemic of tuberculosis. However, the radical cure of pulmonary tuberculosis cannot be completely achieved at present. According to res...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569C07K14/35
CPCG01N33/5695C07K14/35G01N2469/20G01N2333/35
Inventor 毕利军陶生策张先恩朱国峰邓教宇张泓泰侯剑王雅果
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES