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OSBPL1A gene and application thereof in protein encoding

A technology that encodes proteins and genes, applied in the field of bioengineering, can solve problems such as low sensitivity, unclear mechanism of action, false positives, etc., and achieve accurate diagnosis of tuberculosis

Inactive Publication Date: 2017-10-20
SHENZHEN UNIV
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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 has high specificity and is currently the gold standard for diagnosing active tuberculosis. Low sensitivity (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 negative and false positive
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
It has been found that tuberculosis patients can express OSBPL1A, but its mechanism of action in tuberculosis is not clear
There is no report about OSBPL1A gene as a diagnostic marker for tuberculosis

Method used

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  • OSBPL1A gene and application thereof in protein encoding
  • OSBPL1A gene and application thereof in protein encoding
  • OSBPL1A gene and application thereof in protein encoding

Examples

Experimental program
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Effect test

Embodiment 1

[0022] The present embodiment divides the crowd into three groups: the present embodiment divides the crowd into three groups: tuberculosis patients, latently infected people and healthy people (20 cases each), by detecting the OSBPL1A gene in each peripheral blood mononuclear cell (PBMC) mRNA changes, and found that its expression tended to be significantly down-regulated in cured tuberculosis patients.

[0023] In this example, the quantitative RT-PCR method was used to detect the expression change of OSBPL1A gene in each case. Specific steps are as follows:

[0024] Step 1: Preparation of Peripheral Blood Mononuclear Cell (PBMC) Suspension

[0025] 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 equal volume of 1M phosphate buffer ( PBS) to mix well to obtain ...

Embodiment 2

[0052] 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 OSBPL1A gene mRNA in each peripheral blood mononuclear cell (PBMC) by a gene chip, it is found that it is significantly different in tuberculosis patients. There was a clear trend of up-regulated expression.

[0053] In this embodiment, gene chips are used to detect differences in gene expression levels of the OSBPL1A gene in TB, LTBI, and HC in tuberculosis samples, including the following four steps:

[0054] Step 1: Chip Preparation

[0055] At present, glass or silicon wafers are mainly used as carriers for preparing chips, and target genes are arranged on the carrier in sequence as probes by spotting method. Target genes can be divided into genomic DNA and cDNA (or artificially synthesized DNA).

[0056] Step 2: Sample Preparation

[0057] The extraction steps of total RNA in the sample to...

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Abstract

The invention discloses an OSBPL1A gene and application thereof in protein encoding, which are used for preparing a product for judging latent tuberculosis infection and abortive tuberculosis. The product for judging latent tuberculosis infection and abortive tuberculosis is preferably a product for judging the latent tuberculosis infection and abortive tuberculosis through real-time quantitative PCR (polymerase chain reaction) or gene chip detection. Proofed by experiments, the OSBPL1A gen has the advantages that the expression level of the OSBPL1A gene in the blood of a tuberculosis patient is obviously higher than the expression level of the OSBPL1A gene in the blood of health people or latently infected people; the OSBPL1A gene can be used as a specific marker gene for diagnosing the tuberculosis, so that the tuberculosis diagnosis is more accurate and quicker.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to the application of the OSBPL1A gene and its encoded protein. Background technique [0002] Tuberculosis (TB) is a chronic infectious disease caused by Mycobacterium tuberculosis infection. 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 active tuberculosis patients. However, current detection techniques for diagnosing active tuberculosis have serio...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q2600/158
Inventor 陈心春
Owner SHENZHEN UNIV