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MS4A6A gene application

An MS4A6A and gene technology, applied in the field of MS4A6A gene, can solve the problems of no significant increase in sensitivity, low sensitivity, and long time consumption.

Inactive Publication Date: 2013-05-01
THE THIRD PEOPLES HOSPITAL OF SHENZHEN
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  • Summary
  • Abstract
  • 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
3) Antibody detection in immunological testing is considered unsuitable for the diagnosis of tuberculosis by the World Health Organization; cellular immunological testing includes tuberculin skin test (TST) and tuberculosis interferon release test (IGRA), which cannot effectively distinguish patients with active tuberculosis and latent tuberculosis infection, although the latter is significantly more sensitive than other tests in patients with active tuberculosis
There is no report on MS4A6A gene as a diagnostic marker for tuberculosis

Method used

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Examples

Experimental program
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Embodiment 1

[0023] 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 MS4A6A 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.

[0024] In this example, the quantitative RT-PCR method was used to detect the expression changes of MS4A6A gene in each case. Specific steps are as follows:

[0025] Step 1: Preparation of Peripheral Blood Mononuclear Cell (PBMC) Suspension

[0026] Add 5ml of lymphocyte separation medium (Fresenius Kabi NOrge As: 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 ) to mix well to obtain a mixed solution, use a pipette to sl...

Embodiment 2

[0057] In this embodiment, the crowd is divided into three groups: 9 cases of tuberculosis patients, 6 cases of latently infected people and healthy people, by detecting the change of MS4A6A gene mRNA in each peripheral blood mononuclear cell (PBMC) by gene chip, it is found that it is significantly different in tuberculosis patients. There was a clear trend of up-regulated expression.

[0058] In this embodiment, the difference in gene expression level of MS4A6A gene in TB, LTBI, and HC in tuberculosis samples is detected by gene chip, including the following four steps:

[0059] Step 1: Chip Preparation

[0060] At present, the preparation of chips mainly uses glass or silicon wafers as the carrier, and the target genes are arranged on the carrier in sequence as probes by the spotting method. The target genes can be divided into genomic DNA and cDNA (or artificially synthesized DNA).

[0061] Step 2: Sample Preparation

[0062] The extraction steps of total RNA in the samp...

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Abstract

The invention provides an MS4A6A gene application, and relates to the preparation of products to distinguish latent tuberculosis infection and active tuberculosis. The preferred products include the products utilizing real-time quantitative PCR or gene chip detection to distinguish the latent tuberculosis infection and the active tuberculosis. According to the experimental results, the expression of the MS4A6A gene is obviously higher in the blood of tuberculosis patients than in healthy people or latent infection crowds, and therefore, the MS4A6A gene can serve as a special marking gene for the diagnosis of tuberculosis, so as to allow the tuberculosis diagnosis to be more accurate and quicker.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to the application of MS4A6A 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 a...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 陈心春张国良周伯平杨倩婷张明霞蔡毅
Owner THE THIRD PEOPLES HOSPITAL OF SHENZHEN
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