Use of CD157 gene

A CD157-R, gene technology, applied in the field of bioengineering, can solve the problems of long time, high laboratory biosafety requirements, false positives, etc., and achieve the effect of accurate tuberculosis diagnosis

Inactive Publication Date: 2013-04-24
THE THIRD PEOPLES HOSPITAL OF SHENZHEN
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, current detection techniques for diagnosing active tuberculosis have serious deficiencies and cannot meet the requirements of clinical and tuberculosis prevention and control: 1) Sputum Mycobacterium tuberculosis microbiological examination is highly specific and is currently the gold standard for diagnosing active tuberculosis. Low reliability (less than 40%), long time-consuming (1-2 months for tuberculosis culture), and high requirements for laboratory biosafety
2) Mycobacterium tuberculosis gene detection, although the purpose of rapid diagnosis (1 day) has been achieved, the sensitivity of genetic detection directly from sputum samples has not been significantly improved, and there are problems of false negatives and false positives
[0005] There is no report about CD157 gene as a diagnostic marker for tuberculosis

Method used

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Examples

Experimental program
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Embodiment 1

[0024] In this embodiment, the population is divided into three groups: tuberculosis patients, latent infection population and healthy population (20 cases each). By detecting the change of CD157 gene mRNA in each peripheral blood mononuclear cell (PBMC), it is found that it is present in tuberculosis patients. A clear trend of up-regulated expression.

[0025] In this example, quantitative RT-PCR method was used to detect the expression changes of CD157 gene in each case. Specific steps are as follows:

[0026] Step 1: Preparation of Peripheral Blood Mononuclear Cell (PBMC) Suspension

[0027] Add 5ml of lymphocyte separation medium (Fresenius Kabi NOrgeAs: LYS3773) into the centrifuge tube; take 2ml of heparin anticoagulated venous blood from the above-mentioned confirmed tuberculosis patients, patients with latent infection and healthy people and an equivalent amount of 1M phosphate buffer solution (PBS) Mix well to obtain a mixed solution, use a pipette to slowly superi...

Embodiment 2

[0056] In this embodiment, the crowd is divided into three groups: 9 cases of tuberculosis patients, 6 cases of latently infected people and healthy people, and the changes of CD157 gene mRNA in each peripheral blood mononuclear cell (PBMC) were detected by gene chip, and it was found that it was significantly different in tuberculosis patients. There was a clear trend of up-regulated expression.

[0057] In this embodiment, the gene chip is used to detect the difference in gene expression level of CD157 gene in TB, LTBI, and HC in tuberculosis samples, including the following four steps:

[0058] Step 1: Chip Preparation

[0059] At present, glass or silicon wafers are mainly used as carriers to prepare chips, and target genes are arranged in sequence as probes on the carrier by spotting method. Target genes can be divided into genomic DNA and cDNA (or artificially synthesized DNA).

[0060] Step 2: Sample Preparation

[0061] The extraction steps of total RNA in the sample...

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Abstract

The present invention provides a use of a CD157 gene for the preparation of products for distinguishing latent tuberculosis infection and active tuberculosis. The products for distinguishing latent tuberculosis infection and active tuberculosis preferably comprise products adopting real-time quantitative PCR or microarrays to determine and distinguish latent tuberculosis infection and active tuberculosis. Experiments prove that the expression level of the CD157 gene of the present invention in tuberculosis patient blood is significantly higher than that of healthy people as well as people with latent infection, so the CD157 gene can be used as a specific marker gene for diagnosis of tuberculosis, thus making tuberculosis diagnosis more accurate and faster.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to the application of CD157 gene. Background technique [0002] Tuberculosis (TB) is a chronic infectious disease caused by Mycobacterium tuberculosis infection. Mycobacterium tuberculosis can not only cause pulmonary tuberculosis (85%), but also cause tuberculosis in multiple organs outside the lungs. Although there are currently effective anti-tuberculosis drugs, tuberculosis is still the number one killer of infectious diseases, and about 2 million people die from tuberculosis every year in the world; about one-third of the world's population is infected with Mycobacterium tuberculosis, which is called About 10% of people with latent tuberculosis infection (LTBI) will eventually develop active tuberculosis. [0003] Due to the lack of effective tuberculosis vaccine, the prevention and control of tuberculosis mainly depends on early detection, treatment and isolation of ac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 陈心春蔡毅周伯平杨倩婷张明霞
Owner THE THIRD PEOPLES HOSPITAL OF SHENZHEN
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